USPTO Patents Application 09478188

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IN THE UNITED STATES PATENT AND TRADEMARK OFFICE £

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In re Patent Application of

BEN SHEN, WEN LIU, STEVEN D.
CHRISTENSON and SCOTT STANDAGE

For: GENE CLUSTER FOR

PRODUCTION OF THE ENEDIYNE
ANTITUMOR ANTIBIOTIC C-1027

Patent Application

Assistant Commissioner for Patents

Washington, D.C. 20231

By Express Mail No: EL160743652US
Dated: January 5, 2000

PATENT APPLICATION TRANSMITTAL

Sir:

Transmitted herewith for filing is the patent application of inventor(s) Ben Shen, Wen
Liu, Steven D. Christenson and Scott Standage, for "GENE CLUSTER FOR PRODUCTION OF
THE ENEDIYNE ANTITUMOR ANTIBIOTIC C-1027." Enclosed are:

1. 64 pages of the specification, including 71 claims and an abstract.

2. 11 sheets of drawings.

3 . 79 pages of Sequence Listing.

4. An oath or declaration of the inventors (unsigned).

San Francisco, California

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•i— i

The filing fee is being deferred at this time.

Dated: January 5, 2000.

Tom Hunter (Reg. No. 38,498)

MAJESTIC, PARSONS, SIEBERT & HSUE P.C.

Four Embarcadero Center, Suite 1 100

San Francisco, California 941 1 1-4106

Telephone: (415)248-5500

Atty. Docket: 2500.128US1 Facsimile: (415)362-5418

UCRef: 99-174-1

-2-

Docket No: 2500.128US1
Client Ref: 99-174-1

In the United States Patent and Trademark Office
U.S. Patent Application For

GENE CLUSTER FOR PRODUCTION OF THE ENEDIYNE
ANTITUMOR ANTIBIOTIC C-1027

Inventor(s): BEN SHEN, a citizen of the Peoples Republic of China, residing at

1842 Rushmore Lane, Davis, CA 95616, USA

WEN LIU, a citizen of the Peoples Republic of China, residing at the
Institute of Medicinal biotechnology, Tiantan, Beijing, 100005, China

STEVEN D. CHRISTENSON, a citizen of the United States of
America, residing at 1079 Monarch Lane, Davis, CA, 95616, USA

SCOTT STANDAGE, a citizen of the United Kingdom, residing at
63 Tudor Road, Bornet, Herts, EN5 5NW, U.K.

Assignee: The Regents of the University of California

Entity: Small Entity

Majestic, Parsons, Siebert & Hsue P.C.

Four Embarcadero Center, Suite 1100
San Francisco, CA 941 1 1-4106
Tel: 415 248-5500
Fax: 415 362-5418

GENE CLUSTER FOR PRODUCTION OF THE ENEDIYNE
ANTITUMOR ANTIBIOTIC C-1027

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit under 35 U.S.C. §1 19 of provisional
5 application USSN 60/1 15,434, filed on January 6, 1999, which is herein incorporated by
reference in its entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY
SPONSORED RESEARCH AND DEVELOPMENT

This work was supported in part by a grant from the Cancer Research
10 Coordinating Committee, University of California, the National Institutes of Health grant
CA78747, and the Searle Scholars Program/The Chicago Community Trust. The
Government of the United States of America may have certain rights in this invention.

FIELD OF THE INVENTION

This invention relates to the field of enediyne antibiotics. In particular this
15 invention elucidates the gene cluster controlling the biosynthesis of the C-1027 enediyne.

BACKGROUND OF THE INVENTION

The enediyne antibiotics are currently the focus of intense research activity in
the fields of chemistry, biology, and medical sciences, because of their unique molecular
architecture, biological activities, and modes of actions (Doyle and Borders (1995) Enediyne

20 antibiotics as antitumor agents. Marcel-Dekker, New York, Thorson et al. (1999) Bioorg.
Chem., 27: 172-188). Since the unveiling of the structure of neocarzinostatin chromophore
(Edo et al. (1985) Tetrahedron Lett. 26: 331-340) in 1985, the enediyne family has grown
steadily. Thus far, there have been three basic groups within the enediyne antibiotic family:
(a) the calicheamicin/esperamicin type, which includes the calicheamicins, the esperamicins,

25 and namenamicin, (b) the dynemicin type, and (c) the chromoprotein type, consisting of an
apoprotein and an unstable enediyne chromophore. The latter group includes
neocarzinostatin, kedarcidin, C-1027 (Fig. 1), and maduropeptin, whose enediyne
chromophore structures have been established, as well as several others whose enediyne
chromophore structures are yet to be determined due to their instability (Thorson et al.

-1-

(1999) Bioorg. Chem., 27: 172-188). N1999A2, in contrast to the other chromoproteins,
exists as an enediyne chromophore alone despite the fact that its structure is very similar to
the other chromoprotein chromophore (Ando et a/.(1998) Tetra. Letts., 39: 6495-6480).

As a family, the enediyne antibiotics are the most potent, highly active
5 antitumor agents ever discovered. Some members are 1 000 times more potent than
adriamycin, one of the most effective, clinically used antitumor antibiotics (Zhen et al
(1989) J, Antibiot. 42: 1294-1298). All members of this family contain a unit consisting of
two acetylenic groups conjugated to a double bond or incipient double bond within a nine or
ten-membered ring; i.e., the enediyne core as exemplified by C-1027 in Fig. 1. As the

10 consequence of this structural feature, these compounds share a common mechanism of
action: the enediyne core undergoes an electronic rearrangement to form a transient
benzenoid diradical, which is positioned in the minor groove of DNA so as to damage DNA
by abstracting hydrogen atoms from deoxyriboses on both strands (Fig. 1). Reaction of the
resulting deoxyribose carbon-centered radicals with molecular oxygen initiates a process that

15 results in both single-strand and double-strand DNA cleavages (Doyle and Borders (1995)
Enediyne antibiotics as antitumor agents. Marcel-Dekker, New York; Ikemoton et al (1995)
Proc. Natl Acad. Set USA 92:10506-10510; Myers et al (1997) J. Am. Chem. Soc. 1 19:
2965-2972; Stassinopoulos et al (1996) Science 272: 1943-1946; Thorson et al (1999)
Bioorg. Chem., 27: 172-188; Xu et al. (1997) J. Am. Chem. Soc. 119: 1133-1134). This

20 novel mechanism of DNA damage has important implications for their application as potent
cancer chemo therapeutic agents (Doyle and Borders (1995) supra.; Sievers et al (1999)
Blood 93:3678-3684).

As an alternative to making structural analogs of microbial metabolites by
chemical synthesis, manipulations of genes governing secondary metabolism offer a

25 promising alternative allowing preparation of these compounds biosynthetically (Cane et al
(1998) Science 282: 63-68; Hutchinson and Fujii. (1995) Ann. Rev. Microbiol 49: 201-38;
Katz and Donadio (1993) Ann. Rev. Microbiol. 47: 875-912). The success of the latter
approach depends critically on the availability of novel genetic systems and on genes
encoding novel enzyme activities. The enediynes offer a distinct opportunity to study the

30 biosynthesis of their unique molecular scaffolds and the mechanism of self-resistance to
extremely cytotoxic natural products. Elucidation of these aspects provides access to
rational engineering of enediyne biosynthesis for novel drug leads and makes it possible to
construct enediyne overproducing strains by de-regulating the biosynthetic machinery. In

addition, elucidation of an enediyne gene cluster contributes to the general field of
combinatorial biosynthesis by expanding the repertoire of novel polyketide synthase (PKS)
and deoxysugar biosynthesis genes as well as other genes uniquely associated with enediyne
biosynthesis, leading to the making of novel enediynes via combinatorial biosynthesis.

5 SUMMARY OF THE INVENTION

This invention provides nucleic acid sequences and characterization of the
gene cluster responsible for the biosynthesis of the enediyne C-1027 (produced by
Streptomyces globisporus). In particular structural and functional characterization is
provided for the 50 open reading frames (ORFs) comprising this gene cluster. Thus, in one

10 embodiment, this invention provides an isolated nucleic acid comprising a nucleic acid
selected from the group consisting of a nucleic acid encoding any of C-1027 open reading
frames (ORFs) -7 through 42, excluding ORF 9 (cagA), a nucleic acid encoding a
polypeptide encoded by any of C-1027 open reading frames (ORFs) -7 through 42, excluding
ORF 9 (cagA); and a nucleic acid amplified by polymerase chain reaction (PCR) using

15 primer pairs that amplify any of C-1027 open reading frames (ORFs) -7 through 42,

excluding ORF 9 (cagA). In one embodiment, preferred nucleic acids comprise a nucleic
acid encoding at least two (more preferably at least three or more) open reading frames
(ORFs) selected from the group consisting of ORF-1 through ORF 42, excluding ORF 9
(cagA).

20 In another embodiment this invention provides an isolated nucleic acid

comprising a nucleic acid that specifically hybridizes under stringent conditions to an open
reading frame (ORF) of the C-1027 biosynthesis gene cluster, excluding ORF 9 (cagA), and
can substitute for the ORF to which it specifically hybridizes to direct the synthesis of an
enediyne. In certain embodiments this also includes nucleic acids that would stringently

25 hybridizes indicated above, but for, the degeneracy of the nucleic acid code. In other words,
if silent mutations could be made in the subject sequence so that it hybridizes to he indicated
sequence(s) under stringent conditions, it would be included in certain embodiments.
Particularly preferred nucleic acids comprises a nucleic acid that specifically hybridizes
under stringent conditions to a nucleic acid selected from the group consisting of ORF -7,

30 ORF -6, ORF -5, ORF -4, ORF -3, ORF -2, ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4,
ORF 5, ORF 6, ORF 7, ORF 8, ORF 10, ORF 11, ORF 12, ORF 13, ORF 14, ORF 15, ORF
16, ORF 17, ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23, ORF 24, ORF 25, ORF

26, ORF 27, ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33, ORF 34, ORF 35, ORF
36, ORF 37, ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42. Particularly preferred
isolated nucleic acid comprises a nucleic acid selected from the group consisting of ORF -7,
ORF -6, ORF -5, ORF -4, ORF -3, ORF -2, ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4,
5 ORF 5, ORF 6, ORF 7, ORF 8, ORF 10, ORF 11, ORF 12, ORF 13, ORF 14, ORF 15, ORF
16, ORF 17, ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23, ORF 24, ORF 25, ORF
26, ORF 27, ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33, ORF 34, ORF 35, ORF
36, ORF 37, ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42. The nucleic acid may
comprises a nucleic acid that is a single nucleotide polymorphism (SNP) of a nucleic acid

10 selected from the group consisting of ORF -7, ORF -6, ORF -5, ORF -4, ORF -3, ORF -2,
ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 81, ORF 1,
ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 8, ORF 10, ORF 1 1, ORF 12, ORF 13,
ORF 14, ORF 15, ORF 16, ORF 17, ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23,
ORF 24, ORF 25, ORF 26, ORF 27, ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33,

15 ORF 34, ORF 35, ORF 36, ORF 37, ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42.

This invention also provides an isolated gene cluster comprising open reading
frames encoding polypeptides sufficient to direct the assembly of a C-1027 enediyne or a C-
1027 enediyne analogue. The gene cluster may be present in a cell, more preferably in a
bacterial cell (e.g. Actinornycetes, Actinoplanetes, Actinomadura, Micromonospora, or

20 Streptomycetes). Particular preferred bacterial cells include, but are not limited to

Streptomyces globisporus, Streptomyces lividans, Streptomyces coelicolor, Micromonospora
echinospora spp. calichenisis, Actinomadura verrucosopora, Micromonospora chersina,
Streptomyces carzinostaticus, and Actinomycete L585-6. The gene cluster may contain one
or more open reading frames is operatively linked to a heterologous promoter (e.g. a

25 constitutive or an inducible promoter).

This invention also provides for an polypeptide encoded by any one or more
of the nucleic acids described herein.

Also provided are host cell(s) (e.g. eukaryotic cells or bacterial cells as
described herein) transformed with one or more of the expression vectors described herein.

30 Preferred host cells are transformed with an exogenous nucleic acid comprising a gene

cluster encoding polypeptides sufficient to direct the assembly of a C-1027 enediyne or a C-
1027 enediyne analogue. In certain embodiments, heterologous nucleic acid may comprise
only a portion of the gene cluster, but the cell will still be able to express an enediyne.

This invention also provides methods of chemically modifying a biological
molecule. The methods involve contacting a biological molecule that is a substrate for a
polypeptide encoded by a C-1027 biosynthesis gene cluster open reading frame, with a
polypeptide encoded by a C-1027 biosynthesis gene cluster open reading frame whereby the
5 polypeptide chemically modifies the biological molecule. In one preferred embodiment, the
polypeptide is an enzyme selected from the group consisting of a hydroxylase, a
homocysteine synthase, a dNDP-glucose dehydrogenase, a citrate carrier protein, a C-methyl
transferase, an N-methyl transferase, an aminotransferase, a CagA apoprotein, an NDP-
glucose synthase, an epimerase, an acyl transferase, a coenzyme F390 synthase, and

10 epoxidase hydrolase, an anthranilate synthase, a glycosyl transferase, a monooxygenase, a
type II condensation protein, an aminomutase, a type II adenylation protein, an O-methyl
transferase, a P-450 hydroxylase, an oxidoreductase, and a proline oxidase. In a preferred
embodiment the method involves contacting the biological molecule with at least two
(preferably at least three or more) different polypeptides encoded by C-1027 biosynthesis

15 gene cluster open reading frames. The contacting may be in a host cell (e.g. a eukaryotic cell
or a bacterial cell) or the contacting can be ex vivo. The biological molecule can be an
endogenous metabolite produced by said host cell or an exogenous supplied metabolite. In
preferred embodiments, the host cell is a bacterial cell or eukaryotic cell (e.g., a mammalian
cell, a yeast cell, a plant cell, a fungal cell, an insect cell, etc.). In certain preferred

20 embodiments, the host cell synthesizes sugars and glycosylates the biological molecule. In
other preferred embodiments, the host cell synthesizes deoxysugars. The method can further
involve contacting the biological molecule with a polyketide synthase or a non-ribosomal
polypeptide synthetase. The contacting can be in a cell (e.g., a bacterial cell) or ex vivo. In
one preferred embodiment the method comprises contacting the biological molecule with at

25 substantially all of the polypeptides encoded by C-1027 biosynthesis gene cluster open
reading frames and said method produces an enediyne or enediyne analogue. In another
preferred embodiment, the biological molecule is a fatty acid and the biological molecule is
contacted with a C-1027 orf polyeptide selected from the group consisting of an epoxide
hydrase, a monooxygenase, an iron-sulfer flavoprotein, a p-450 hydroxylase, an

30 oxidoreductase, and a proline oxidase. In certain embodiments, the biological molecule is a
fatty acid and said biological molecule is contacted with a plurality of C-1027 orf
polypeptides comprising an epoxide hydrase, a monooxygenase, an iron-sulfer flavoprotein,
a p-450 hydroxylase, an oxidoreductase, and a proline oxidase. In one especially preferred

embodiment ,the biological molecule is contacted with polypeptides encoded by ORF 17,
ORF20, ORF21, ORF29, ORF30, ORF32, ORF35, and ORF38, In another especially
preferred embodiment, the biological molecule is contacted with polypeptides encoded by
ORF 15, ORF 16, ORF 28, ORF3, ORF 14, and ORF 13, and, in certain embodiments, ORF
5 4 and ORF 3 as well.

In certain embodiments, the method may comprise contacting a sugar with
one or more C-1027 open reading frame polypeptides selected from the group consisting of a
dNDP-glucose synthase, a dNDP glucose dehydratase, an epimerase, an aminotransferase, a
C-methyltransferase, an N-methyltransferase, and a glycosyl transferase. Particularly

10 preferred variant of this method comprise contacting a dNDP-glucose with a plurality of C-
1027 open reading frame polypeptides comprising a dNDP-glucose synthase, a dNDP
glucose dehydratase, an epimerase, an aminotransferase, a C-methyltransferase, an N-
methyltransferase, and a glycosyl transferase.

In certain other embodiments, the method comprises contacting an amino acid

15 with one or one or more C-1027 open reading frame polypeptides selected from the group

consisting of a hydroxylase, an aminomutase, a type II NRPS condensation enzyme, a type II
NRPS adenylation enzyme, and a type II peptidyl carrier protein. These methods may
involve contacting an amino acid with a plurality of C-1027 open reading frame polypeptides
comprising a hydroxylase, a halogenase, an aminomutase, a type II NRPS condensation

20 enzyme, a type II NRPS adenylation enzyme, and a type II peptidyl carrier protein. In
particularly preferred embodiments, the amino acid is a tyrosine.

This invention also provides a method of synthesizing a chromaprotein type
enediyne core, said method comprising contacting a fatty acid with one or more C-1027 orf
polypeptides selected from the group consisting of an epoxide hydrase, a monooxygenase, an

25 iron-sulfer flavoprotein, a p-450 hydroxylase, an oxidoreductase, and a proline oxidase. In
preferred embodiments, the fatty acid may be contacted with a plurality of C-1027 orf
polypeptides comprising an epoxide hydrase, a monooxygenase, an iron-sulfer flavoprotein,
a p-450 hydroxylase, an oxidoreductase, and a proline oxidase. In particularly preferred
embodiments, the fatty acid is contacted with polypeptides encoded by ORF 17, ORF20,

30 ORF21, ORF29, ORF30, ORF32, ORF35, and ORF38.

In still yet another embodiment, this invention provides a method of
synthesizing a deoxysugar. This method involves contacting a sugar with one or more C-
1027 open reading frame polypeptides selected from the group consisting of a dNDP-glucose

synthase, a dNDP glucose dehydratase, an epimerase, an aminotransferase, a C-
methyl transferase, an N-methyltransferase, and a glycosyl transferase. In preferred
embodiments, this method involves contacting a dNDP-glucose with a plurality of C-1027
open reading frame polypeptides comprising a dNDP-glucose synthase, a dNDP glucose
5 dehydratase, an epimerase, an aminotransferase, a C-methyltransferase, an N-

methyltransferase, and a glycosyl transferase. In particularly preferred embodiments, the
dNDP-glucose is contacted with polypeptides encoded by ORF17, ORF20, ORF21, ORF29,
ORF30, ORP32, ORF35, and ORF38.

This invention also provides methods of synthesizing a beta amino acid by

10 contacting an amino acid with one or one or more C-1027 open reading frame polypeptides
selected from the group consisting of a hydroxylase, an aminomutase, a type II NRPS
condensation enzyme, a type II NRPS adenylation enzyme, and a type II peptidyl carrier
protein. The method preferably comprises contacting an amino acid with a plurality of C-
1027 open reading frame polypeptides comprising a hydroxylase, a halogenase, an

15 aminomutase, a type II NRPS condensation enzyme, a type II NRPS adenylation enzyme,
and a type II peptidyl carrier protein. Particularly preferred embodiments comprise
contacting the amino acid (e.g. tyrosine) with polypeptides encoded by ORF 4, ORF1 1,
ORF24, ORF23, ORF25, and ORF26.

Also provided are methods of synthesizing an enediyne or an enediyne

20 analogue. These methods involve culturing a cell (e.g. a eukaryotic cell or a bacterium)
comprising a recombinantly modified C-1027 gene cluster under conditions whereby said
cell expresses said enediyne or enediyne analogue; and recovering the enediyne or enediyne
analogue. In preferred embodiments, the gene cluster is present in a bacterium (e.g.,
Actinomycetes, Actinoplanetes, Actinomadura, Micromonospora^ or Streptomycetes).

25 Particularly preferred bacteria include, but are not limited to Streptomyces globisporus,
Streptomyces lividans, Streptomyces coelicolor, Micromonospora echinospora spp.
calichenisis, Actinomadura verrucosopora, Micromonospora chersina, Streptomyces
carzinostaticus, and Actinomycete L585-6. In another preferred embodiment, the gene
cluster is present in a eukaryotic cell (e.g. a mammalian cell, a yeast cell, a plant cell, a

30 fungal cell, an insect cell, etc.). The host cell can be one that synthesizes sugars and
glycosylates the enediyne or enediyne analogue. The host can be one that synthesizes
deoxy sugars.

-7-

This invention also provides a method of making a cell (e.g., a bacterial or
eukaryotic cell) resistant to an enediyne or an enediyne metabolite. This method involves
expressing in the cell one or more isolated C-1027 open reading frame nucleic acids that
encode a protein selected from the group consisting of a CagA apoprotein, a SgcB
5 transmembrane efflux protein, a transmembrane transport protein, a Na+/H+ transporter, an
ABC transport, a glycerol phosphate tranporter, and a UvrA-like protein. In preferred
embodiments, the isolated C-1027 open reading frame nucleic acids are selected from the
group consisting of ORF 9, ORF2, ORF 27, ORF 0, ORF 1 c-terminus, ORF 2, and ORF 1
N-terminus. Certain embodiments exclude cagA (ORF 9).
10 In one embodiment, this invention specifically excludes one or more of open

reading frames -7 through 42. In particular, in one embodiment this invention excludes cagA
(ORF 9), and/or sgcA (ORF 1), and/or sgcB (ORF 2).

DEFINITIONS

The terms "C-1027 open reading frame", and "C-1027 ORF" refer to an open
15 reading frame in the C-1027 biosynthesis gene cluster as isolated from Streptomyces
globisporus. The term also embraces the same open reading frames as present in other
enediyne-synthesizing organisms (e.g. other strains and/or species of Streptomyces,
Actinomyces, and the like). The term encompasses allelic variants and single nucleotide
polymorphisms (SNPs). In certain instances the C-1027 ORF is used synonymously with the
20 polypeptide encoded by the C-1027 ORF and may include conservative substitutions in that
polypeptide. The particular usage will be clear from context.

The terms "isolated" "purified" or "biologically pure" refer to material which
is substantially or essentially free from components which normally accompany it as found
in its native state. With respect to nucleic acids and/or polypeptides the term can refer to
25 nucleic acids or polypeptides that are no longer flanked by the sequences typically flanking
them in nature.

The terms "polypeptide", "peptide" and "protein" are used interchangeably
herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers
in which one or more amino acid residue is an artificial chemical analogue of a
30 corresponding naturally occurring amino acid, as well as to naturally occurring amino acid
polymers. The term also includes variants on the traditional peptide linkage joining the
amino acids making up the polypeptide.

The terms "nucleic acid" or "oligonucleotide" or grammatical equivalents
herein refer to at least two nucleotides covalently linked together. A nucleic acid of the
present invention is preferably single-stranded or double stranded and will generally contain
phosphodiester bonds, although in some cases, as outlined below, nucleic acid analogs are
5 included that may have alternate backbones, comprising, for example, phosphoramide
(Beaucage et al (1993) Tetrahedron 49:1925) and references therein; Letsinger (1970) J.
Org. Chem. 35:3800; Sprinzl et al. (1977) Eur. J. Biochem. 81: 579; Letsinger et al (1986)
Nucl Acids Res. 14: 3487; Sawai et al (1984) Chem. Lett. 805, Letsinger et al (1988) J. Am.
Chem. Soc. 110: 4470; and Pauwels et al (1986) Chemica Scripta 26: 141 9),

10 phosphorothioate (Mag et al. (1991) Nucleic Acids Res. 19: 1437; and U.S. Patent No.
5,644,048), phosphorodithioate (Briu et al (1989) J. Am. Chem. Soc. 1 1 1 :2321, O-
methylphophoroamidite linkages (see Eckstein, Oligonucleotides and Analogues: A
Practical Approach, Oxford University Press), and peptide nucleic acid backbones and
linkages (see Egholm (1992) J. Am. Chem. Soc. 1 14:1895; Meier et al (1992) Chem. Int. Ed.

15 Engl 31: 1008; Nielsen (1993) Nature, 365: 566; Carlsson et al (1996) Nature 380: 207).
Other analog nucleic acids include those with positive backbones (Denpcy et al (1995)
Proc. Natl Acad. Sci. USA 92: 6097; non-ionic backbones (U.S. Patent Nos. 5,386,023,
5,637,684, 5,602,240, 5,216,141 and 4,469,863; Angew. (1991) Chem. Ml Ed. English 30:
423; Letsinger et al (1988) J. Am. Chem. Soc. 1 10:4470; Letsinger et al (1994) Nucleoside

20 & Nucleotide 13: 1597; Chapters 2 and 3, ASC Symposium Series 580, "Carbohydrate

Modifications in Antisense Research", Ed. Y.S. Sanghui and P. Dan Cook; Mesmaeker et al
(1994), Bioorganic & Medicinal Chem. Lett. 4: 395; Jeffs et al (1994) J. Biomolecular NMR
34:17; Tetrahedron Lett. 37:743 (1996) and non-ribose backbones, including those described
in U.S. Patent Nos. 5,235,033 and 5,034,506, and Chapters 6 and 7, ASC Symposium Series

25 580, Carbohydrate Modifications in Antisense Research, Ed. Y.S. Sanghui and P. Dan Cook,
Nucleic acids containing one or more carbocyclic sugars are also included within the
definition of nucleic acids (see Jenkins et al (1995), Chem. Soc. Rev. pp 169- 176). Several
nucleic acid analogs are described in Rawls, C & E News June 2, 1997 page 35. These
modifications of the ribose-phosphate backbone may be done to facilitate the addition of

30 additional moieties such as labels, or to increase the stability and half-life of such molecules
in physiological environments.

The term "heterologous" as it relates to nucleic acid sequences such as coding
sequences and control sequences, denotes sequences that are not normally associated with a

region of a recombinant construct, and/or are not normally associated with a particular cell.
Thus, a "heterologous" region of a nucleic acid construct is an identifiable segment of
nucleic acid within or attached to another nucleic acid molecule that is not found in
association with the other molecule in nature. For example, a heterologous region of a
5 construct could include a coding sequence flanked by sequences not found in association
with the coding sequence in nature. Another example of a heterologous coding sequence is a
construct where the coding sequence itself is not found in nature (e.g., synthetic sequences
having codons different from the native gene). Similarly, a host cell transformed with a
construct which is not normally present in the host cell would be considered heterologous for

10 purposes of this invention.

A "coding sequence" or a sequence which "encodes" a particular polypeptide
(e.g. a PKS, an NRPS, etc.), is a nucleic acid sequence which is ultimately transcribed and/or
translated into that polypeptide in vitro and/or in vivo when placed under the control of
appropriate regulatory sequences. In certain embodiments, the boundaries of the coding

15 sequence are determined by a start codon at the 5 r (amino) terminus and a translation stop
codon at the 3 1 (carboxy) terminus. A coding sequence can include, but is not limited to,
cDNA from procaryotic or eucaryotic mRNA, genomic DNA sequences from procaryotic or
eucaryotic DNA, and even synthetic DNA sequences. In preferred embodiments, a
transcription termination sequence will usually be located 3' to the coding sequence.

20 Expression "control sequences" refers collectively to promoter sequences,

ribosome binding sites, polyadenylation signals, transcription termination sequences,
upstream regulatory domains, enhancers, and the like, which collectively provide for the
transcription and translation of a coding sequence in a host cell. Not all of these control
sequences need always be present in a recombinant vector so long as the desired gene is

25 capable of being transcribed and translated.

"Recombination" refers to the reassortment of sections of DNA or RNA
sequences between two DNA or RNA molecules. "Homologous recombination" occurs
between two DNA molecules which hybridize by virtue of homologous or complementary
nucleotide sequences present in each DNA molecule.

30 The terms "stringent conditions" or "hybridization under stringent conditions"

refers to conditions under which a probe will hybridize preferentially to its target

subsequence, and to a lesser extent to, or not at all to, other sequences. "Stringent

hybridization" and "stringent hybridization wash conditions" in the context of nucleic acid

-10-

hybridization experiments such as Southern and northern hybridizations are sequence
dependent, and are different under different environmental parameters. An extensive guide
to the hybridization of nucleic acids is found in Tijssen (1993) Laboratory Techniques in
Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes part I chapter
5 2 Overview of principles of hybridization and the strategy of nucleic acid probe assays^
Elsevier, New York. Generally, highly stringent hybridization and wash conditions are
selected to be about 5°C lower than the thermal melting point (T m ) for the specific sequence
at a defined ionic strength and pH. The T m is the temperature (under defined ionic strength
and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Very

10 stringent conditions are selected to be equal to the T m for a particular probe.

An example of stringent hybridization conditions for hybridization of
complementary nucleic acids which have more than 100 complementary residues on a filter
in a Southern or northern blot is 50% formamide with 1 mg of heparin at 42°C, with the
hybridization being carried out overnight. An example of highly stringent wash conditions is

15 0.15 M NaCl at 72°C for about 15 minutes. An example of stringent wash conditions is a
0.2x SSC wash at 65°C for 15 minutes (see, Sambrook et al (1989) Molecular Cloning - A
Laboratory Manual (2nd ed.) Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor
Press, NY, for a description of SSC buffer). Often, a high stringency wash is preceded by a
low stringency wash to remove background probe signal. An example medium stringency

20 wash for a duplex of, e.g., more than 100 nucleotides, is lx SSC at 45°C for 15 minutes. An
example low stringency wash for a duplex of, e.g,, more than 100 nucleotides, is 4-6x SSC at
40°C for 15 minutes. In general, a signal to noise ratio of 2x (or higher) than that observed
for an unrelated probe in the particular hybridization assay indicates detection of a specific
hybridization. Nucleic acids which do not hybridize to each other under stringent conditions

25 are still substantially identical if the polypeptides which they encode are substantially
identical. This occurs, e.g., when a copy of a nucleic acid is created using the maximum
codon degeneracy permitted by the genetic code.

Expression vectors are defined herein as nucleic acid sequences that are direct
the transcription of cloned copies of genes/cDNAs and/or the translation of their mRNAs in

30 an appropriate host. Such vectors can be used to express genes or cDNAs in a variety of

hosts such as bacteria, bluegreen algae, plant cells, insect cells and animal cells. Expression

vectors include, but are not limited to, cloning vectors, modified cloning vectors, specifically

designed plasmids or viruses. Specifically designed vectors allow the shuttling of DNA

-11-

between hosts, such as bacteria-yeast or bacteria- animal cells. An appropriately constructed
expression vector preferably contains: an origin of replication for autonomous replication in
a host cell, a selectable marker, optionally one or more restriction enzyme sites, optionally
one or more constitutive or inducible promoters. In preferred embodiments, an expression
5 vector is a replicable DNA construct in which a DNA sequence encoding a one or more PKS
and/or NRPS domains and/or modules is operably linked to suitable control sequences
capable of effecting the expression of the products of these synthase and/or synthetases in a
suitable host. Control sequences include a transcriptional promoter, an optional operator
sequence to control transcription and sequences which control the termination of

10 transcription and translation, and so forth.

The term "conservative substitution" is used in reference to proteins or
peptides to reflect amino acid substitutions that do not substantially alter the activity
(specificity or binding affinity) of the molecule. Typically conservative amino acid
substitutions involve substitution one amino acid for another amino acid with similar

15 chemical properties (e.g. charge or hydrophobicity). The following six groups each contain
amino acids that are typical conservative substitutions for one another: 1) Alanine (A),
Serine (S), Threonine (T); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N),
Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M),
Valine (V); and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

20 The "group consisting of ORF-1 through ORF 42" refers to the group

consisting of ORF -7, ORF -6, ORF -5, ORF -4, ORF -3, ORF -2, ORF -1, ORF 0, ORF 1,
ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 81, ORF 1, ORF 2, ORF 3, ORF 4,
ORF 5, ORF 6, ORF 7, ORF 8, ORF 9, ORF 10, ORF 11, ORF 12, ORF 13, ORF 14, ORF
15, ORF 16, ORF 17, ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23, ORF 24, ORF

25 25, ORF 26, ORF 27, ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33, ORF 34, ORF
35, ORF 36, ORF 37, ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42 as identified in
Tables I and II. In certain embodiments ORF 9 (cagA) is excluded.

A "biological molecule that is a substrate for a polypeptide encoded by a
enediyne (e.g., C-1027) biosynthesis gene" refers to a molecule that is chemically modified

30 by one or more polypeptides encoded by open reading frame(s) of the C-1027 biosynthesis
gene cluster. The "substrate" may be a native molecule that typically participates in the
biosynthesis of an enediyne, or can be any other molecule that can be similarly acted upon
by the polypeptide.

-12-

A "polymorphism" is a variation in the DNA sequence of some members of a
species. A polymorphism is thus said to be "allelic/' in that, due to the existence of the
polymorphism, some members of a species may have the unmutated sequence (i.e. the
original "allele") whereas other members may have a mutated sequence {i.e. the variant or
5 mutant "allele"). In the simplest case, only one mutated sequence may exist, and the
polymorphism is said to be diallelic. In the case of diallelic diploid organisms, three
genotypes are possible. They can be homozygous for one allele, homozygous for the other
allele or heterozygous. In the case of diallelic haploid organisms, they can have one allele or
the other, thus only two genotypes are possible. The occurrence of alternative mutations can

10 give rise to trialleleic, etc. polymorphisms. An allele may be referred to by the nucleotide(s)
that comprise the mutation.

"Single nucleotide polymorphism" or "SNPs are defined by their
characteristic attributes. A central attribute of such a polymorphism is that it contains a
polymorphic site, "X," most preferably occupied by a single nucleotide, which is the site of

15 the polymorphism's variation (Goelet and Knapp U.S. patent application Ser. No.

08/145,145). Methods of identifying SNPs are well known to those of skill in the art (see,
e.g., U.S. Patent 5,952,174).

Abbreviations used herein include LB, Luria-Bertani; NGDH, dNDP-glucose
4,6-dehydratase ; nt, nucleotide; ORF, open reading frame; PCR, polymerase chain reaction;

20 PEG, polyethyleneglycol; PKS, polyketide synthase; RBS, ribosomal binding site; Apr,
apramycin; R, resistant; Th, thiostrepton; WT, wild-type; and TS, temperature sensitive

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 illustrates the structures of C-1027 chromophore and the benzenoid
diradical intermediate proposed to initiate DNA cleavage.
25 Figure 2 illustrates a scheme using C-1027 open reading frame polypeptides

for the synthesis of deoxysugars.

Figure 3 A illustrates a scheme using C-1027 open reading frame polypeptides
for the synthesis of a p-amino acid.

Figure 3B illustrates a scheme using C-1027 open reading frame polypeptides
30 for the synthesis of a benzoxazolinate.

-13-

Figure 4 illustrates the synthesis of the enediyne core and final assembly of

the C-1027 enediyne.

Figures 5 A, 5B, and 5C illustrate the organization of the C-1027 enediyne
biosynthetic gene cluster. Figure 5A shows a restriction map of the 75-kb sgc gene cluster
5 from S. globisporus as represented by three cosmid clones. Figure 5B illustrates the genetic
organization of the sgcA, sgcB, and cagA genes, showing that they are clustered in the sgc
gene cluster. Probe 1, the 0.55-kb dNDP-glucose 4,6-dehydratase gene fragment from
pBS1002. Probe 2, the 0.73-kb cagA fragment from pBS1003. A, Apal; B, BamHl; E,
EcoRl; K, Kpnl, S, Sacll; Sp, Sphl. Figure 5C shows the genetic organization of the C-1027
10 biosynthesis gene cluster.

Figure 6 shows the DNA and deduced amino acid sequences of the 3,0-kb
BamHl fragment from pBS1007, showing the sgcA and sgcB genes. Possible RBSs are
boxed. The presumed translational start and stop sites are in boldface. Restriction enzyme
sites of interest are underlined. The amino acids, according to which the degenerated PCR
15 primer were designed for amplifying the dNDP-glucose 4,6-dehydratase gene from S.
globisporus, are underlined.

Figure 7 shows the amino acid sequence alignment of SgcA with three other
dNDP-glucose 4,6-dehydratases. Gdh, TDP-glucose 4,6-dehydratase of S. erythraea
(AAA6821 1); MtmE, TDP-glucose 4,6-dehydratase in the mithramycin pathway of S.
20 argillaceus (CAA7 1 847); TylA2, TDP-glucose 4,6-dehydratase in the tylosin pathway of S,
fradiae (S49054). Given in parentheses are protein accession numbers. The apa fold with
the NAD + -binding motif of GxGxxG is boxed.

Figures 8 A and 8B show disruption of sgc A by single crossover homologous
recombination. Figure 8 A shows construction of sgcA disruption mutant and restriction
25 maps of the wild-type S. globisporus C-1027 and S. globisporus SB 1001 mutant strains
showing predicted fragment sizes upon BamHl digestion. Figures 8B and 8C show a
Southern analysis of S. globisporus C-1027 (lane 1) and S. globisporus SB1001 (lanes 2, 3,
and 4, three individual isolates) genomic DNA, digested with BamHl, using (Figure 8B)
pOJ260 vector or (Figure 8C) the 0.75-kb SaclllKpnl fragment of sgcA from pBS1012 as a
30 probe, respectively. B, BamHl; K, Kpnl; S, Sacll.

Figures 9 A, 9B, 9B, and 9D illustrate the determination of C-1027 production
in various S. globisporus strains by assaying their antibacterial activity against M luteus.

-14-

Figure 9A:1, S. globisporusC-1027; 2,3, and 4, S. globisporus SB 1001 (three individual
isolates); 5, S. globisporus AF67; 6, S. globisporus AF40. Figure 9B: 1, S. globisporus C-
1027; 2, S. globisporus SB1001 (pWHM3); 3 and 4, S. globisporus SB1001 (pBS1015) (two
individual isolates). Both S. globisporus SB 1001 (pWHM3) and S. globisporus SB 1001
5 (pBS1015) were grown in the presence of 5 jig/mL thiostrepton. Figure 9C: 1, S.

globisporusC-1027; 2, S. globisporus SB1001 (pBS1015); 3. S. globisporus SB1001; 4. S.
globisporus SB1001 (pWHM3); 5. S. globisporus AF40; 6. 5. globisporus AF44. All 5.
globisporus strains were grown in the absence of thiostrepton. Figure 9D: 1.5. globisporus
(pKC1139); 2. S. globisporus (pBS1018).

1 0 DETAILED DESCRIPTION

This invention provides a complete gene cluster regulating the biosynthesis of
C-1027, the most potent member of the enediyne antitumor antibiotic family. C-1027 is
produced by Streptomyces globisporus C-1027 and consists of an apoprotein (encoded by the
cagA gene) and a non-peptidic chromophore. The C-1027 chromophore could be viewed as
15 being derived biosynthetically from a benzoxazolinate, a deoxyamino hexose, a p-amino
acid, and an enediyne core. Adopting a strategy to clone the C-1027 biosynthesis gene
cluster by mapping a putative dNDP-glucose 4,6-dehydratase (NGDH) gene to cagA, we
localized 75 kb contiguous DNA from S. globisporus encoding a complete C-1027 gene
cluster.

20 Initial sequencing of the cloned gene cluster revealed two genes, sgcA and

sgcB } that encode an NGDH enzyme and a transmembrane efflux protein, respectively, and
confirmed that the cagA gene resides approximately 14 kb upstream of the sgcA,B locus.
The involvement of the cloned gene cluster in C-1027 biosynthesis was demonstrated by
disrupting the sgcA gene to generate C-1027-nonproducing mutants and by complementing

25 the sgcA mutants in vivo to restore C-1027 production.

Subsequent DNA sequence analysis provided the complete enediyne C-1027
gene cluster sequence (SEQ ID NOs: 1 and 2) revealing 50 open reading frames which are
summarized in Tables I and II. These results represent the first cloning of a gene cluster for
enediyne anti-tumor antibiotic biosynthesis.

30

-15-

Table L Summary of the C-1027 gene cluster open reading frames. Table 1. C-1027 gene
cluster open reading frames (-7 to 26), primers for ORF amplification, and proposed
functions

orf# Size Relative Primers Function Seq

position ID

No.

orf -

648

658-11

Fwd:

ATG

GGC

ATG

ACG

GGT

very weak

3

(-7)

bp

Rev:

CTA

GAG

GAT

CCC

GGG

homology to

4

ny U.X. uAy labc

orf -

549

147 8 -

Fwd :

Alb

Lbb

Al 1

bbb

vlldl

c

(-6)

bp

93 0

Rev :

rpp 71

1 LA

bL 1

bl b

HAT
bAl

uJ.b-

niLcbbi viuy

potentiator

>~\ "V" f~y I - d "1 T1
pi D L. fc= -LI1

orf-

1065

2713-

Fwd:

ATG

ACC

ATC

GCC

ACT

N-truncated

7

C-5)

bp

1649

Rev:

TCA

GAG

GCC

GAG

CAC

Methionine

8

o t m "I - Vi^aoci (Hi \r o 1 \7

oyiiuiiciofc; v J- -J- e -L y

psuedogene)

orf-

387

3238-

Fwd:

ATG

AGC

TCG

CTA

CTG

Viral

9

(-4)

bp

2851

Rev :

CTA

GGA

GCC

GGT

ppp
LbL

transcription

1 Pi
J. U

factor

orf-

1530

4971-

Fwd:

ATG

AGC

AGC

AGC

GCC

Viral Homo log

11

(-3)

bp

3442

Rev :

TCA

TTC

GTC

GGC

TGC

possibly primase

JLZ

orf-

3027

5982-

Fwd:

GTG

AGG

GCT

CTG

CCG

Glycerol-

13

(-2)

bp

7478

Rev :

TCA

bAL

ppp

pp7\

bbA

ppp

bbb

irnospnaue a±5v_

1 A

Transporter

\ OI1UA VJ.-L. U.y

IcblbL dllO C /

orf -

2328

99 00 -

Fwd :

GTG

AGC

GTC

7\ pn

ALU

p 7\ p

bAb

uviA-iiKe Qiug

(-1)

bp

7573

Rev :

TCA

ACC

CGC

CCT

GCG

resistance pump

j. b

orf -

13 68

11349-

Fwd :

Alb

Abb

7\ rpp

Alb

Lib

prpp

bib

TVTa + of flnv

in a /xi em ux.

1 7
J. /

0

bp

9982

Rev :

GTG

GCT

GTG

CTC

p p A

bLA

pump

T Q

JL O

orf -

99 9

28590-

Fwd :

Al b

a rr*
Abb

A

Alb

Lib

prpp

bib

s3 "NTT 1 n_/f1 1 1 pnao
CUM lr yiULUbc

1 Q

1

bp

29588

Rev :

rpP "A

ppp
bLL

P7\ p

bAL

ppp
bbb

prpp

b 1 L

^ £i Vlt V~ ^1 "t~ ^ O £2l

Qcnyurduabc

ort -

1566

z y 6 6 z -

Fwd :

bib

A. H7±
ALA

ULA

bib

± J_ Ctiio LiLCLLUlJX. O.HG

21

2

bp

31197

Rev :

TCA

TGT

bbL

ppp
Lbb

rprpp

1 lb

sin ux protein

orf-

1311

31280-

Fwd:

GTG

GAG

TAC

TGG

AAC

Coenzyme F3 90

23

3

bp

32590

Rev:

TCA

GGC

CTG

AGG

GGC

synthase

24

phenylacetyl-CoA

ligase

orf -

1584

32809-

Fwd:

GTG

CCC

CAC

GGT

GCA

phenol

25

4

bp

34392

Rev:

CTA

CAG

CCC

TCC

GAG

hydroxylase

26

chlorophenol-4 -

monooxygena s e

orf -

bp

35274-

Fwd:

ATG

TCT

TCA

ACC

CGT

citrate

27

5

34458

Rev:

TCA

GCC

GCG

CAG

GAA

transport

28

protein

orf -

1272

17924-

Fwd:

ATG

CTG

GAG

AAA

TGC

C -methyl

29

6

16653

Rev:

TCA

GAC

GAG

CTC

CTT

transferase

30

hydroxylase

orf-

735

16653-

Fwd:

ATG

GAG

TAC

GGC

CCC

N-

31

-16-

7

bp

15919

Rev :

TCA

TGC

CGT

GCG

CAC

methyltransf eras

32

orf -

1233

15922-

Fwd:

ATG

AGC

GGC

GGC

CCG

e

Aminotransferase

33

8

bp

14690

Rev :

TCA

CCT

CGC

CGG

ACG

34

orf -

432

14643-

Fwd:

ATG

TCG

TTA

CGT

CAC

CagA

35

9

bp

14212

Rev:

TCA

GCC

GAA

GGT

CAG

36

orf-

1068

13012-

Fwd:

ATG

AAG

GCA

CTT

GTA

dNTP-glucose

37

10

bp

14079

Rev:

TCA

GGC

CGC

GAT

CTC

synthase

38

orf-

1485

12835-

Fwd:

GTG

GAC

GTG

TCA

GCG

Hydroxylase,

39

11

bp

11351

Rev :

TCA

GGA

CCG

CGC

ACC

Halogenase

4 U

orf -

579

25564-

Fwd:

ATG

AAG

CCG

ATC

GGG

dNTP-4-keto-6-

41

12

bp

24986

Rev:

TCAGGA CGA CTT GTT

deoxyglucose

42

3 , 5-epimerase

orf-

1137

24702-

Fwd:

ATG

CCT

TCC

CCC

TTC

3-0-

43

13

bp

23566

Rev:

TCA

GGT

GCG

CTC

GGC

acyl transferase

44

orf -

1455

22878-

Fwd:

GTG

AGA

GAC

GGC

CGG

Coenzyme F-390

45

14

bp

21424

Rev:

TCA CGT GGT GAT GGC

Synthase

46

Phenylacetyl CoA

Ligase

orf-

1482

21407-

Fwd:

ATG

ACC

GAC

CAG

TGC

Anthranilate

47

15

bp

19926

Rev:

TPS

CAA

CTC

Synthase I

48

orf -

663

19929-

Fwd:

GTG

AGC

TTG

TGG

TCT

Anthranilate

49

16

bp

19267

Rev:

TCA

GGC

CGG

TTC

GGC

Synthase II

50

orf-

1161

19191-

Fwd:

GTG

CGT

CCC

TTC

CGT

epoxide

51

17

bp

18031

Rev:

TCA

GCG

GAG

CGG

ACG

hydrolase

52

orf -

423

35938-

Fwd:

ATG

CCA

GCA

CCG

ACT

Unknown

53

18

bp

35516

Rev:

TCA

GTC

GTT

GCC

GCG

54

orf-

1380

27214-

Fwd:

ATG

CGG

GTG

ATG

ATC

glycosyl

55

19

bp

28593

Rev:

TCA

TCG

GTC

CGC

CTC

transferase

56

orf -

1356

25815-

Fwd:

ATG

ACC

AAG

CAC

GCC

squalene

57

20

bp

27170

Rev:

TCA

TAC

GGC

GGC

GCC

monooxygenase

58

orf -

672

23546-

Fwd:

GTG

AGC

GCA

CAA

CTC

hypothetical Fe-

59

21

bp

22875

Rev:

TCA

CGG

CTG

TGC

CTG

S flavoprotein

60

orf -

816

35274-

Fwd:

ATG

TCT

TCA

ACC

CGT

haloacetate

61

22

bp

34458

Rev:

TCA

GCC

GCG

CAG

GAA

dehalogenase

62

hydrolase

orf -

1380

37559-

Fwd :

ATG

ACG

ACG

TCC

GAC

peptide

63

23

bp

38938

Rev :

TCA

GGA

GGT

GAA

GGG

synthetase

64

orf-

1620

40986-

Fwd:

ATG

GCA

TTG

ACT

CAA

Histidine

65

24

bp

39367

Rev:

TCA

GCG

CAG

CTG

GAT

Ammonia lyase

66

orf -

1560

42611-

Fwd:

ATG

ACG

CGG

CCG

GTG

Type II

67

25

bp

41052

Rev:

TCA GCG GGT GAG CCG

adenylation

68

protein

orf -

282

38983-

Fwd:

GTG

TCC

ACC

GTT

TCC

Type II peptidyl

69

26

bp

39264

Rev:

TCA

CTG

CGT

TCC

GGA

carrier protein

70

-17-

Table II. C-1027 gene cluster open reading frames (27 to 42), primers for ORF
amplification, and proposed functions

ORF Relative Primers Function SEQ

Position ID

NO.

orf -

43945

-46023

Fwd:

GTG

TGC

CCG

GTG

ACA

GAC

Antibiotic

71

27

Rev:

TCA

GCC

CAC

GGG

CTG

GGA

Transporter

72

orf-

46167

-47171

Fwd:

GTG

TTG

GGC

GAT

GAG

GAC

0-

73

28

Rev:

TCA

GAC

CGC

GGA

CAT

CTG

methyl transfer

74

ase

orf - -

4 72 2 7

-484 85

Fwd :

ATG

GCC

GGC

CTG

GTC

ATG

p450

75

O Q

Rev :

TCA

GGA

CCC

GAG

GGT

CAC

hvdroxvlase

76

nr f _

KJ X- _L

48610

-4 97 14

Fwd :

GTG

GAC

CAG

ACG

TCT

ACG

Oxidoreduct ase

77

3 0

Rev :

TCA

TGC

AGG

TGC

AGC

GTG

78

orf —

J \J O ~J \J

Fwd :

ATG

AGG

CCG

CTC

GTT

CGG

Unknown

79

Rev :

TCA TCC CGG CCC GGC GGC

Protein

80

5142 0

-52341

Fwd :

ATG

AGA

ACG

CGG

CGA

CGC

Oxidoreduct ase

81

32

Rev:

TCA

CGG

CCG

GAG

GCG

TAC

82

orf -

53241

-54074

Fwd :

GTG

TAT

CAG

CCG

GAC

TGT

Unknown

83

33

Rev:

CTA

CTC

ATT

CCA

GTT

GTG

Protein

84

orf -

54230

-55379

Fwd:

ATG

TCT

ACG

GGC

TAT

CTC

Unknown

85

34

Rev:

TCA

GCC

GCC

GGT

GGC

GCC

Protein

86

orf -

56027

-56881

Fwd:

ATG

TTC

TCC

CCC

GCC

GCC

Oxidase/

87

35

Rev:

TCA

GTA

CGC

CTG

GTG

GGC

Dehydrogenase

88

orf-

56928

-57730

Fwd:

ATG

AAT

TCG

CTC

GAC

GAC

Unknown

89

36

Rev

: TCA GCT CCC GGT CGC CGC

Protein

90

orf -

57834

-58304

Fwd:

ATG

ACC

GCG

ACG

AAT

CCT

Regulatory

91

37

Rev:

CTA

GGC

GGC

GCG

TCC

CGC

92

orf-

58440

-60091

Fwd:

ATG

AGC

ACC

ACG

GCC

GAG

Oxidoreduct ase

93

38

Rev:

TCA

GCC

GCG

CGC

CGA

CGG

94

orf-

60092

-60622

Fwd:

ATG

ACC

CTG

GAG

GCC

TAC

Regulatory

95

39

Rev:

TCA

TGC

GGG

GCT

CCC

GGT

96

orf-

60940

-62020

Fwd:

GTG

AAA

AGT

GAC

TCT

GCC

Regulatory

97

40

Rev:

TCA

ACG

GCG

AGT

TGG

CTG

98

orf-

62045

-62899

Fwd:

GTG

ACC

ACG

AAC

ACC

ATC

Regulatory

99

41

Rev:

TCA

CCC

GCG

ATC

TCG

ATC

10

orf -

62788

-63164

Fwd :

(partial ORF)

p450

10

42

Rev:

TCA

CCT

CGC

CGT

ACT

CAC

hydroxylase

10

5

Surprisingly, sequence analysis failed to reveal any gene that resembles a
polyketide synthase. The C-1027 open reading frames, however, encode polypeptides
exhibiting a wide variety of enzymatic activities {e.g., epoxide hydrase, monooxygenase,
oxidoreductase, P-450 hydroxylase, etc.). The isolated C-1027 gene cluster can be used to

-18-

synthesize C-1027 enediyne antibiotics and/or analogues thereof. The C-1027 gene cluster
can be modified and/or augmented to increase C-1027 and/or C-1027 analogue production.

Alternatively, various components of the C-1027 gene cluster can be used to
synthesize and/or chemically modify a wide variety of metabolites. Thus, for example, ORF
5 6 (C-methyltransferase) can be used to methylate a carbon, while ORF 12, an epimerase, can
be used to change the conformation of a sugar. The ORFs can be combined in their native
configuration or in modified configurations to synthesize a wide variety of
biomolecules/metabolites. Thus, for example, various combinations of C-1027 open reading
frames can be used to synthesize an enediyne core, to synthesize a deoxy sugar, to synthesize

10 a p-amino acid, to make a benzoxazolinate, etc (see, e.g., Figures 2, 3, and 4).

The native C-1027 gene cluster ORFs can be re-ordered, modified, and
combined with other biosynthetic units (e.g. polyketide synthases (PKSs) or catalytic
domains thereof and/or non-ribosomal polypeptide synthetases (NRPSs) or catalytic domains
thereof) to produce a wide variety of molecules. Large chemical libraries can be produced

15 and then screened for a desired activity.

The C-1027 gene cluster also includes a number of drug resistance genes (see,
e.g., Table 2) that confer resistance to C-1027 and/or metabolites involved in C-1027
biosynthesis thereby permitting the cell to complete the enediyne biosynthesis. These
resistance genes can be used to confer enediyne resistance on a cell lacking such resistance

20 or to augment the enediyne resistance of a cell that does tolerate enediynes. Such cells can
be used to produce high levels of enediynes and/or enediyne metabolites, and/or enediyne
analogues.

Table III. C-1027 cluster drug resistance genes.

ORF

Protein

Mechanism

ORF 9:

CagA apoprotein

Drug sequestering

ORF 2:

SgcB transmembrane efflux protein

Drug exporting

ORF 27

Transmembrane transport protein

Drug exporting

ORF0

Na + /H + transporter

Drug exporting

ORF-1

ABC transport (C- terminus)

Drug exporting

ORF -2

Glycerol phosphate transporter

Drug exporting

ORF-1

UvrA-like protein (N-terminus)

DNA repairing

-19-

L Isolation, preparation, and expression of C-1027 nucleic acids.

The C-1027 gene cluster nucleic acids can be isolated, optionally modified,
and inserted into a host cell to create and/or modify a metabolic (biosynthetic) pathway and
5 thereby enable that host cell to synthesize and/or modify various metabolites. Alternatively
the C-1027 gene cluster nucleic acids can be expressed in the host cell and the encoded C-
1027 polypeptide(s) recovered for use as chemical reagents, e.g. in the ex vivo synthesis
and/or chemical modification of various metabolites. Either application typically entails
insertion of one or more nucleic acids encoding one or more isolated and/or modified C-1027

10 enediyne open reading frames in a suitable host cell. The nucleic acid(s) are typically in an
expression vector, a construct containing control elements suitable to direct expression of the
C-1027 polypeptides. The expressed C-1027 polypeptides in the host cell then act as
components of a metabolic/biosynthetic pathway (in which case the synthetic product of the
pathway is typically recovered) or the C-1027 polypeptides themselves are recovered. Using

15 the sequence information provided herein, cloning and expression of C-1027 nucleic acids
can be accomplished using routine and well known methods.

A) C-1027 nucleic acids.

The nucleic acids comprising the C-1027 gene cluster are identified in Tables
I and are listed in the sequence listing provided herein. In particular, Table 1 identifies genes

20 and functions of open reading frames (ORFs) in the C-1027 enediyne biosynthesis gene
cluster and identifies primers suitable for the amplification/isolation of any one or more of
the C-1027 open reading frames. Of course, using the sequence information provided herein,
other primers suitable for amplification/isolation of one or more C-1027 open reading frames
can be determined according to standard methods well known to those of skill in the art (e.g.

25 using Vector NTI Suite™, InforMax, Gaithersberg, MD, USA).

Typically such amplifications will utilize the DNA or RNA of an organism
containing the requisite genes (e.g. Streptomyces globisporus) as a template. Typical
amplification conditions include the following PCR temperature program: initial denaturing
at 94°C for 5 min, 24-36 cycles of 45 sec at 94°C, 1 min at 60°C, 2 min at 72°C, followed by

30 additional 7 min at 72°C. One of skill will appreciate that optimization of such a protocol,
e.g. to improve yield, etc. is routine (see, e.g., U.S. Patent No. 4,683,202; Innis (1990) PCR

-20-

Protocols A Guide to Methods and Applications Academic Press Inc. San Diego, CA, etc).
In addition, primer may be designed to introduce restriction sites and so facilitate cloning of
the amplified sequence into a vector.

In one embodiment, this invention provides nucleic acids for the recombinant
5 expression of an enediyne (e.g. a C-1027 enediyne or an analogue thereof). Such nucleic
acids include isolated gene cluster(s) comprising open reading frames encoding polypeptides
sufficient to direct the assembly of the enediyne. In other embodiments of this invention, the
C-1027 open reading frames may be unchanged, but the control elements (e.g. promoters,
enhancers, etc.) may be modified. In still other embodiments, the nucleic acids may encode

10 selected components (e.g. one or more C-1027 or modified C-1027 open reading frames)
and/or may optionally contain other heterologous biosynthetic elements including, but not
limited to polyketide synthase (PKS) and/or non-ribosomal polypeptide synthetase (NRPS)
modules or enzymatic domains.

Such variations may be introduced by design, for example to modify a known

15 molecule in a specific way, e.g. by replacing a single substituent of the enediyne with
another, thereby creating a derivative enediyne molecule of predicted structure.
Alternatively, variations can be made randomly, for example by making a library of
molecular variants of a known enediyne by systematically or haphazardly replacing one or
open reading frames in the biosynthetic pathway. Production of alternative/modified

20 enediyne, and hybrid enediyne PKSs and/or NRPSs and hybrid systems is described below.

Using the information provided herein other approaches to cloning the desired
sequences will be apparent to those of skill in the art. For example, the enediyne, and/or
optionally PKS and/or NRPS modules or enzymatic domains of interest can be obtained
from an organism that expresses such, using recombinant methods, such as by screening

25 cDNA or genomic libraries, derived from cells expressing the gene, or by deriving the gene
from a vector known to include the same. The gene can then be isolated and combined with
other desired biosynthetic elements using standard techniques. If the gene in question is
already present in a suitable expression vector, it can be combined in situ, with, e.g., other
PKS subunits, as desired. The gene of interest can also be produced synthetically, rather

30 than cloned. The nucleotide sequence can be designed with the appropriate codons for the

particular amino acid sequence desired. In general, one will select preferred codons for the

intended host in which the sequence will be expressed. The complete sequence can be

assembled from overlapping oligonucleotides prepared by standard methods and assembled

-21-

into a complete coding sequence (see, e.g., Edge (1981) Nature 292:756; Nambair et al
(1984) Science 223: 1299; Jay et al (1984) J. Biol Chem. 259:6311). In addition, it is noted
that custom gene synthesis is commercially available (see, e.g. Operon Technologies,
Alameda, CA).

5 Examples of such techniques and instructions sufficient to direct persons of

skill through many cloning exercises are found in Berger and Kimmel (1989) Guide to
Molecular Cloning Techniques, Methods in Enzymology 152 Academic Press, Inc., San
Diego, CA (Berger); Sambrook et al (1989) Molecular Cloning - A Laboratory Manual (2nd
ed.) Vol 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor Press, NY; Ausubel (19
1 0 1 994) Current Protocols in Molecular Biology, Current Protocols, a joint venture between
Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., U.S. Patent 5,017,478; and
European Patent No. 0,246,864.

Expression of f C-1027 open reading frames.

The choice of expression vector depends on the sequence(s) that are to be
15 expressed. Any transducible cloning vector can be used as a cloning vector for the nucleic
acid constructs of this invention. However, where large clusters are to be expressed, it
phagemids, cosmids, Pis, YACs, BACs, PACs, HACs or similar cloning vectors be used for
cloning the nucleotide sequences into the host cell. Phagemids, cosmids, and BACs, for
example, are advantageous vectors due to the ability to insert and stably propagate therein
20 larger fragments of DNA than in M13 phage and lambda phage, respectively. Phagemids
which will find use in this method generally include hybrids between plasmids and
filamentous phage cloning vehicles. Cosmids which will find use in this method generally
include lambda phage-based vectors into which cos sites have been inserted. Recipient pool
cloning vectors can be any suitable plasmid. The cloning vectors into which pools of
25 mutants are inserted may be identical or may be constructed to harbor and express different
genetic markers (see, e.g., Sambrook et a/., supra). The utility of employing such vectors
having different marker genes may be exploited to facilitate a determination of successful
transduction.

In preferred embodiments of this invention, vectors are used to introduce C-
30 1027 biosynthesis genes or gene clusters into host (e.g. Streptomyces) cells. Numerous
vectors for use in particular host cells are well known to those of skill in the art. For
example described in Malpartida and Hopwook, (1984) Nature, 309:462-464; Kao et al,

-22-

(1994), Science, 265: 509-512; and Hopwood et al, (1987) Methods Enzymol, 153:1 16-166
all describe vectors for use in various Streptomyces hosts.

In one preferred embodiment, Streptomyces vectors are used that include
sequences that allow their introduction and maintenance in E. coli. Such Streptomyces! E.
5 coli shuttle vectors have been described (see, for example, Vara et al, (1989) J. Bacteriol,
171:5872-5881; Guilfoile & Hutchinson (1991) Proc. Natl Acad. Set USA, 88: 8553-8557.)

The wildtype and/or modified C-1027 enediyne open reading frame(s) of this
invention, can be inserted into one or more expression vectors, using methods known to
those of skill in the art. Expression vectors will include control sequences operably linked to

10 the desired open reading frame. Suitable expression systems for use with the present
invention include systems that function in eucaryotic and/or prokaryotic host cells.
However, as explained above, prokaryotic systems are preferred, and in particular, systems
compatible with Streptomyces spp. are of particular interest. Control elements for use in
such systems include promoters, optionally containing operator sequences, and ribosome

15 binding sites. Particularly useful promoters include control sequences derived from
enediyne, and/or PKS, and/or NRPS gene clusters. Other promoters {e.g. ermE* as
illustrated in Example 1) are also suitable. Other bacterial promoters, such as those derived
from sugar metabolizing enzymes, such as galactose, lactose (lac) and maltose, will also find
use in the present constructs. Additional examples include promoter sequences derived from

20 biosynthetic enzymes such as tryptophan (trp), the beta -lactamase (bid) promoter system,

bacteriophage lambda PL, and T5. In addition, synthetic promoters, such as the tac promoter
(U.S. Patent 4,551,433), which do not occur in nature also function in bacterial host cells. In
Streptomyces, numerous promoters have been described including constitutive promoters,
such as ErmE and TcmG (Shen and Hutchinson, (1994) J. Biol Chem. 269: 30726-30733),

25 as well as controllable promoters such as actl and actlll (Pleper et al, (1995) Nature, vol.
378: 263-266; Pieper et al, (1995) Am, Chem. Soc, 1 17: 1 1373-1 1374; and Wiesmann et
al, (1995) Chem. & Biol 2: 583-589).

Other regulatory sequences may also be desirable which allow for regulation
of expression of the enediyne open reading frame(s) relative to the growth of the host cell.

30 Regulatory sequences are known to those of skill in the art, and examples include those

which cause the expression of a gene to be turned on or off in response to a chemical or

physical stimulus, including the presence of a regulatory compound. Other types of

regulatory elements may also be present in the vector, for example, enhancer sequences.

-23-

Selectable markers can also be included in the recombinant expression
vectors. A variety of markers are known which are useful in selecting for transformed cell
lines and generally comprise a gene whose expression confers a selectable phenotype on
transformed cells when the cells are grown in an appropriate selective medium. Such
5 markers include, for example, genes that confer antibiotic resistance or sensitivity to the
plasmid.

The various enediyne cluster open reading frames, and/or PKS, and/or NRPS
clusters or subunits of interest can be cloned into one or more recombinant vectors as
individual cassettes, with separate control elements, or under the control of, e.g., a single

10 promoter. The various open reading frames can include flanking restriction sites to allow for
the easy deletion and insertion of other open reading frames so that hybrid synthetic
pathways can be generated. The design of such unique restriction sites is known to those of
skill in the art and can be accomplished using the techniques described above, such as site-
directed mutagenesis and PCR.

15 Methods of cloning and expressing large nucleic acids such as gene clusters,

including PKS- or NRPS-encoding gene clusters, in cells including Streptomyces are well
known to those of skill in the art (see, e.g., Stutzman-Engwall and Hutchinson (1989) Proc.
Natl Acad. Set USA, 86: 3135-3139; Motamedi and Hutchinson (1987) Proc. Natl Acad.
Scl USA, 84: 4445-4449; Grim et al (1994) Gene, 151: 1-10; Kao et al (1994) Science,

20 265: 509-512; and Hopwood et al (1987) Meth. Enzymol, 153: 1 16-166). In some
examples, nucleic acid sequences of well over lOOkb have been introduced into cells,
including prokaryotic cells, using vector-based methods (see, for example, Osoegawa et al,
(1998) Genomics, 52: 1-8; Woon et al, (1998) Genomics, 50: 306-316; Huang et al, (1996)
Nucl Acids Res., 24: 4202-4209). In addition, the cloning and expression of C-1027

25 enediyne is illustrated in Example 1 .

Q Host cells.

The vectors described above can be used to express various protein
components of the enediyne, and/or enediyne shunt metabolites, and/or other modified
metabolites for subsequent isolation and/or to provide a biological synthesis of one or more
30 desired biomolecules (e.g. C-1027 and/or a C-1027 analogue, etc.). Where one or more
proteins of the enediyne biosynthetic gene cluster are expressed (e.g. overexpressed) for
subsequent isolation and/or characterization, the proteins are expressed in any prokaryotic or

-24-

eukaryotic cell suitable for protein expression. In one preferred embodiment, the proteins

are expressed in E. coll

Host cells for the recombinant production of the subject enediynes, enediyne

metabolites, shunt metabolites, etc. can be derived from any organism with the capability of
5 harboring a recombinant enediyne gene cluster and/or subset thereof. Thus, the host cells of

the present invention can be derived from either prokaryotic or eucaryotic organisms.

Preferred host cells are those of species or strains {e.g. bacterial strains) that naturally

express enediynes. Such host cells include, but are not limited to Actinomycetes,

Actinoplanetes, and Streptomycetes, Actinomadura, Micromonospra, and the like.
10 Particularly preferred host cells include, but are not limited to Streptomyces globisporus,

Streptomyces lividans, Streptomyces coelicolor, Micromonospora echinospora spp.

calichenisis, Actinomadura verrucosopora, Micromonospora chersina, Streptomyces

carzinostaticus, and Actinomycete L585-6. Other suitable host cells include, but are not

limited to S. verticillis S. arnbofaciens, S. avermitilis, S. azureus, S. cinnamonensis, S.
15 coelicolor, S. curacoi, S. erythraeus, S.fradiae, S. galilaeus, S. glaucescens, S.

hygroscopicus, S. lividans, S. parvulus, S. peucetius, S. rimosus, S. roseofulvus, S.

thermotolerans, and S. violaceoruber {see, e.g., Hopwood and Sherman (1990) Ann. Rev.

Genet. 24: 37-66; O'Hagan (1991) The Polyketide Metabolites, Ellis Horwood Limited,

etc.).

20 In certain embodiments, a eukaryotic host cell is preferred {e.g. where certain

glycosylation patterns are desired). Suitable eukaryotic host cells are well known to those of
skill in the art. Such eukaryotic cells include, but are not limited to yeast cells, insect cells,
plant cells, fungal cells, and various mammalian cells (e.g. COS, CHO HeLa cells lines and
various myeloma cell lines).

25 D) Recovery of the expression product.

Recovery of the expression product {e.g., enediyne, enediyne analogue,
enediyne biosynthetic pathway polypeptide, etc.) is accomplished according to standard
methods well known to those of skill in the art. Thus, for example where enediyne
biosynthetic gene cluster proteins are to be expressed and isolated, the proteins can be
30 expressed with a convenient tag to facilitate isolation {e.g. a His 6 ) tag. Other standard

protein purification techniques are suitable and well known to those of skill in the art {see,

-25-

e.g., (Quadri et al (1998) Biochemistry 37: 1585-1595; Nakano et al. (1992) Mol. Gen.
Genet. 232: 313-321, etc.).

Similarly where components (e.g. enediyne biosynthetic cluster orfs) are used
to synthesize and/or modify various biomolecules (e.g. enediynes, enediyne analogues, shunt
5 metabolites, etc.) the desired product and/or shunt metabolite(s) are isolated according to
standard methods well know to those of skill in the art (see, e.g., Carreras and Khosla (1998)
Biochemistry 37: 2084-2088, Deutscher (1990) Methods in Enzymology Volume 182: Guide
to Protein Purification, M. Deutscher, ed. etc.).

IL Use of C-1027 open reading frames in directed biosynthesis.

10 Elements (e.g. open reading frames) of the C-1027 biosynthetic gene cluster

and/or variants thereof can be used in a wide variety of "directed" biosynthetic processes (i.e.
where the process is designed to modify and/or synthesize one or more particular preselected
metabolite(s)). Essentially the entire C-1027 gene cluster can be used to synthesize a C-1027
enediyne and/or a C-1027 enediyne analogue. Individual C-1027 cluster open reading

15 frames can be used to perform chemically modifications on particular substrates and/or to
synthesize various metabolites. Thus, for example, ORF 6 (C-methyltransferase can be used
to methylate a carbon), while ORF 7 (N-methyltransferase) can be used to methylate a
nitrogen. ORF 12, and epimerase, can be used to change the conformation of a sugar, and
ORF 8 (an amino transferase) can be used to aminate a suitable substrate. Similarly,

20 combinations of C-1027 open reading frames can be used to direct the synthesis of various
metabolites (e.g. p-amino acids, deoxysugars, benzoxazolinates, and the like). These
examples, are merely illustrative. One of skill in the art, utilizing the information provided
here, can perform literally countless chemical modifications and/or syntheses using either
"native" enediyne biosynthesis metabolites as the substrate molecule, or other molecules

25 capable of acting as substrates for the particular enzymes in question. Other substrates can
be identified by routine screening. Methods of screening enzymes for specific activity
against particular substrates are well known to those of skill in the art.

The biosyntheses can be performed in vivo, e.g. by providing a host cell
comprising the desired C-1027 gene cluster open reading frames and/or in vivo, e.g., by

30 providing the polypeptides encoded by the C-1027 gene cluster ORFs and the appropriate
substrates and/or cofactors.

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A) Synthesis of enedivnes and enediyne analogues.

In one embodiment, this invention provides for the synthesis of C- 1027
enediynes and/or C-1027 analogues or derivatives. In a preferred embodiment, this is
accomplished by providing a cell comprising a C-1027 gene cluster and culturing the cell
5 under conditions whereby the desired enediyne or enediyne analogue is synthesized. The
cell can be a cell that does not normally synthesize an enediyne and the entire gene cluster
can be transfected into the cell Alternatively, a cell that typically synthesizes enediynes can
be utilized and all or part of the C-1027 gene cluster can be introduced into the cell.

Enediyne derivatives/analogues can be produced by varying the order of, or
10 kind of, gene cluster subunits present in the cell, and/or by changing the host cell (e.g. to a
eukaryotic cell that glycosylates the biosynthetic product), and/or by providing altered
metabolites (e.g. adding exogenous aglycones to a host that carries a gene cassette of the
deoxysugar biosynthesis and glycosylation genes for the production of glycosylated
metabolites), etc.

1 5 In certain embodiments, the host cell need not be transfected with an entire C-

1027 gene cluster. Rather, various components of a C-1027 gene cluster can be altered
within a cell already harboring a C-1027 cluster. By varying or adding various biosynthetic
open reading frames, C-1027 enediyne variants can be produced.

The use of standard techniques of molecular biology (gene disruption, gene

20 replacement, gene supplement) can be used to modulate and/or otherwise alter enediyne
and/or other metabolite (e.g. shunt metabolite) production in an organism that naturally
synthesizes an enediyne (e.g. S. globisporus) or an organism that is modified to synthesize an
enediyne.

In addition, or alternatively, control sequences that alter the expression of
25 various open reading frames can be introduced that alter the amount and/or timing of

enediyne production. Thus, for example, by placing particular C-1027 open reading frames
under control of a constitutive promoter (ermE*) C-1027 production was increased by as
much as 4-fold {see, e.g. Table 3 and Example 1).

30 Table 3. Alteration of C-1027 production by engineering the C-1027 biosynthesis gene
cluster.

Strain Yield (%)

-27-

WT 100

WT/pKC1139 100

WT/erm£*/ORF 2 >150

WT/ORF 9 >100

WT/e7?n£*/ORF 9 <10

WT/ORF 10, 11 >100

WT/ermE*/ORF 10, 1 1 >1 00

WT/ORF 9, 10, 11 >400

ORF2: transmembrane eflux protein; ORF 9: CagA apoprotein; ORF 10: TDP-glucose
synthase; ORF 11; Hydroxylase/halogenase

Where enediyne analogues are synthesized, it will often prove desirable to
5 assay them for biological activity. Such assays are well know to those of skill in the art.
One such assay is illustrated in Example L Briefly, this example depicts an assay of
antibacterial activity against M. luteus as described by Hu et al (1988) J. Antibiot 41: 1575-
1579). Other suitable assays for enediyne activity will be known to those of skill in the art.

B) Use of C-1027 open reading frames to synthesize an enediyne core.

10 The C-1027 open reading frames described herein, or variants thereof, can be

used to synthesize an enediyene core, e.g., from a fatty acid precursor. One such synthetic
pathway is illustrated in Figure 4. This reaction scheme utilizes ORF 17 (epoxide hydrase),
ORF 20 (monooxygenase), ORF 21 (iron-sulfur flavoprotein), ORF 29 (P-450 hydroxylase,
ORF 30 (oxidoreductase), ORF 32 (oxidoreductase), ORF 35 (proline oxidase), and ORF 38

15 (P-450 hydroxylase) to synthesize anenediyne core.

This synthetic pathway, is not considered limiting, but merely illustrative.
Using this as a model, one of ordinary skill in the art can design numerous other synthetic
schemes to produce enediyne cores and/or core variants.

d Use of C-1027 open reading frames to synthesize deoxy sugars.

20 The biosynthesis of various deoxy sugars {e.g., deoxyhexoses) typically share

a common key intermediate -4-keto-6-deoxyglucose nucleoside diphosphate or its analogs,

whose formation from glucose nucleoside diphosphate is catalyzed by the NGDH enzyme,

an NAD + -dependent oxidoreductase (Liu and Thorson (1994) Ann. Rev. Microbiol. 48: 223-

256; Piepersberg (1997) pp. 81-163. In Biotechnology of antibiotics, 2nd ed. W. R. Strohl

-28-

(ed). Marcel Dekker, New York.). Similarly, the C-1027 gene cluster includes an NAGDH
enzyme which can be exploited to synthesize a variety of deoxy sugars.

One illustrative synthetic pathway is shown in Figure 2. This biosynthetic
scheme utilizes ORF 10 (dNDP-glucose synthase), ORF 1 (dNDP-glucose dehydratase),
5 ORF 12 (epimerase), ORF 8 (aminotransferase), ORF 6 (C-methyltransferase), ORF 7 (N-
methyltransferase) and ORF 19 (glycosyl transferase).

This synthetic pathway, is not considered limiting, but merely illustrative.
Using this as a model, one of ordinary skill in the art can design numerous other synthetic
schemes to produce various deoxy sugars.

10 D) Use of C-1027 open reading frames to synthesize B-amino acids.

In still another embodiment, C-1027 biosynthetic polypeptides can be used in
the biosynthesis of p-amino acids. One illustrative synthetic pathway is shown in Figure 3 A.
This biosynthetic scheme utilizes ORF 4 (hydroxylase), ORF 1 1 (hydroxylase/halogenase),
ORF 24 (aminomutase), ORF 23 (type II NRPS condensation enzyme), ORF 25 (type II
1 5 NRPS adenylation enzyme), and ORF 26 (type II peptidyl carrier protein).

Again, this synthetic pathway, is not considered limiting, but merely
illustrative. Using this as a model, one of ordinary skill in the art can design numerous other
synthetic schemes to produce other beta amino acids.

E) Use of C-1027 open reading frames to synthesize benzoxazolinates.

20 The C-1027 open reading frames can also be used to synthesize a

benzoxazolinate. One illustrative synthetic pathway is shown in Figure 3B. This
biosynthetic scheme utilizes ORF 15 (anthranilate synthase I, ORF 16 (anthranilate synthase
II), ORF 4 (phenol hydroxylase/chlorophenol-4-monooxygenase), ORF 11
(Hydroxylase/Halogenase), ORF 28 (O-methyltransferase), ORF 3 (coenzyme F390

25 synthetase, ORF 14 (coenzyme F390 synthetase), and ORF 13 (O-acyltransferase). Again,
this synthetic pathway, is not considered limiting, but merely illustrative. Using this as a
model, one of ordinary skill in the art can design numerous other synthetic schemes to
produce other beta amino acids.

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III. Generation of chemical diversity.

In addition to the directed modification and/or biosynthesis of various
metabolites as described above, the C-1027 biosynthetic gene cluster open reading frames
can be utilized, by themselves or in combination with other biosynthetic subunits (e.g. NRPS
5 and/or PKS modules and/or enzymatic domains of other PKS and/or NRPS systems) to
produce a wide variety of compounds including, but not limited to various enediyne or
enediyne derivatives, various polyketides, polypeptides, polyketide/polypeptide hybrids,
various thiazoles, various sugars, various methylated polypeptides/polyketides, and the like.

As with the directed production of various metabolites described above, such
10 compounds can be produced, in vivo or in vitro, by catalytic biosynthesis, e.g., using large,
enediyne cluster units and/or modular PKSs, NRPSs, and hybrid PKS/NRPS systems. In a
preferred embodiment large combinatorial libraries of cells harboring various
megasynthetases can be produced by the random or directed modification of particular
pathways and then selected for the production of a molecule or molecules of interest. It will
15 be appreciated that, in certain embodiments, such libraries of megasynthetases/modified
pathways, can be used to generate large, complex combinatorial libraries of compounds
which themselves can be screened for a desired activity.

Such combinatorial libraries can be created by the deliberate
modification/variation of selected biosynthetic pathways and/or by random/haphazard
20 modification of such pathways.

A) Directed eneineerine of novel synthetic pathways.

In numerous embodiments of this invention, novel polyketides, polypeptides,
and combinations thereof are created by modifying the entediyne gene cluster ORFs and/or
known PKSs, and/or NRPSs so as to introduce variations into metabolites synthesized by the

25 enzymes. Such variations may be introduced by design, for example to modify a known

molecule in a specific way, e.g. by replacing a single monomeric unit within a polymer with
another, thereby creating a derivative molecule of predicted structure. Such variations can
also be made by adding one or more modules or enzymatic domains to a known PKS or
NRPS or enediyne cluster, or by removing one or more module from a known PKS or

30 NRPS.

Using any of these methods, it is possible to introduce PKS domains, NRPS

domains, and entediyne domains into a megasynthetase. Mutations can be made to the

-30-

native enediyne, and/or NRPS, and/or PKS subunit sequences and such mutants used in
place of the native sequence, so long as the mutants are able to function with other subunits
(domains) in the synthetic pathway. Such mutations can be made to the native sequences
using conventional techniques such as by preparing synthetic oligonucleotides including the
5 mutations and inserting the mutated sequence into the gene encoding a NRPS and/or PKS
subunit using restriction endonuclease digestion, {see, e.g., Kunkel, (1985) Proc. Natl. Acad.
Sci. USA 82: 448; Geisselsoder et al (1987) BioTechniques 5: 786). Alternatively, the
mutations can be effected using a mismatched primer (generally 10-20 nucleotides in length)
which hybridizes to the native nucleotide sequence (generally cDNA corresponding to the

10 RNA sequence), at a temperature below the melting temperature of the mismatched duplex.
The primer can be made specific by keeping primer length and base composition within
relatively narrow limits and by keeping the mutant base centrally located (Zoller and Smith
(1983) Meth, Enzymol. 100: 468). Primer extension is effected using DNA polymerase, the
product cloned and clones containing the mutated DNA, derived by segregation of the primer

15 extended strand, selected. Selection can be accomplished using the mutant primer as a
hybridization probe. The technique is also applicable for generating multiple point
mutations (see, e.g., Dalbie-McFarland et al (1982) Proc. Natl Acad. Sci USA 79:6409).
PCR mutagenesis will also find use for effecting the desired mutations.

J$) Random modification of enediyne pathways.

20 In another embodiment, variations can be made randomly, for example by

making a library of molecular variants (e.g. of a known enediyne) by randomly mutating one
or more elements of the subject gene cluster or by randomly replacing one or more open
reading frames in a gene cluster with one or more of alternative open reading frames.

The various open reading frames can be combined into a single multi-modular

25 enzyme, thereby dramatically increasing the number of possible combinations obtained using
these methods. These combinations can be made using standard recombinant or nucleic acid
amplification methods, for example by shuffling nucleic acid sequences encoding various
modules or enzymatic domains to create novel arrangements of the sequences, analogous to
DNA shuffling methods described in Crameri et al (1998) Nature 391: 288-291, and in U.S.

30 Patents 5,605,793 and in 5,837,458. In addition, novel combinations can be made in vitro,
for example by combinatorial synthetic methods. Novel molecules or molecule libraries, can
be screened for any specific activity using standard methods.

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Random mutagenesis of the nucleotide sequences obtained as described above
can be accomplished by several different techniques known in the art, such as by altering
sequences within restriction endonuclease sites, inserting an oligonucleotide linker randomly
into a plasmid, by irradiation with X-rays or ultraviolet light, by incorporating incorrect
5 nucleotides during in vitro DNA synthesis, by error-prone PCR mutagenesis, by preparing
synthetic mutants or by damaging plasmid DNA in vitro with chemicals. Chemical mutagens
include, for example, sodium bisulfite, nitrous acid, hydroxylamine, agents which damage or
remove bases thereby preventing normal base-pairing such as hydrazine or formic acid,
analogues of nucleotide precursors such as nitrosoguanidine, 5-bromouracil, 2-aminopurine,

10 or acridine intercalating agents such as proflavine, acriflavine, quinacrine, and the like.

Generally, plasmid DNA or DNA fragments are treated with chemicals, transformed into E.
coli and propagated as a pool or library of mutant plasmids.

Large populations of random enzyme variants can be constructed in vivo
using "recombination-enhanced mutagenesis." This method employs two or more pools of,

15 for example, 10 6 mutants each of the wild-type encoding nucleotide sequence that are

generated using any convenient mutagenesis technique, described more fully above, and then
inserted into cloning vectors.

O Incorporation and/or modification of non-C-1027 cluster elements.

In either the directed or random approaches, nucleic acids encoding novel
20 combinations of gene cluster ORFs are introduced into a cell. In one embodiment, nucleic
acids encoding one or more enediyne synthetic cluster ORFS and/or PKS and/or NRPS
domains are introduced into a cell so as to replace one or more domains of an endogenous
gene cluster within a cell. Endogenous gene replacement can be accomplished using
standard methods, such as homologous recombination. Nucleic acids encoding an entire
25 enediyne, enediyne ORF, PKS, NRPS, or combination thereof can also be introduced into a
cell so as to enable the cell to produce the novel enzyme, and, consequently, synthesize the
novel polymer. In a preferred embodiment, such nucleic acids are introduced into the cell
optionally along with a number of additional genes, together called a 'gene cluster,' that
influence the expression of the genes, survival of the expressing cells, etc. In a particularly
30 preferred embodiment, such cells do not have any other enediyne and/or PKS- and/or NRPS-
encoding genes or gene clusters, thereby allowing the straightforward isolation of the
molecule(s) synthesized by the genes introduced into the cell.

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Furthermore, the recombinant vector(s) can include genes from a single
enediyne and/or PKS and/or NRPS gene cluster, or may comprise hybrid replacement PKS
gene clusters with, e.g., a gene for one cluster replaced by the corresponding gene from
another gene cluster. For example, it has been found that ACPs are readily interchangeable
5 among different synthases without an effect on product structure. Furthermore, a given KR
can recognize and reduce polyketide chains of different chain lengths. Accordingly, these
genes are freely interchangeable in the constructs described herein. Thus, the replacement
clusters of the present invention can be derived from any combination of PKS and/or NRPS
gene sets that ultimately function to produce an identifiable polyketide.

10 Examples of hybrid replacement clusters include, but are not limited to,

clusters with genes derived from two or more of the act gene cluster, the whiE gene cluster,
frenolicin (freri), granaticin (gra), tetracenomycin (tern), 6-methylsalicylic acid (6-msas),
oxytetracycline (otc) 9 tetracycline (tet), erythromycin (ery\ griseusin (gris) 9 nanaomycin,
medermycin, daunorubicin, tylosin, carbomycin, spiramycin, avermectin, monensin,

15 nonactin, curamycin, rifamycin and candicidin synthase gene clusters, among others. (For a
discussion of various PKSs, see, e.g., Hopwood and Sherman (1990) Ann. Rev. Genet. 24:
37-66; O'Hagan (1991) The Polyketide Metabolites, Ellis Horwood Limited.

A number of hybrid gene clusters have been constructed, having components
derived from the actjren, tern, gris and gra gene clusters (see, e.g., U.S. Patent 5,712,146).

20 Other hybrid gene clusters, as described above, can easily be produced and screened using
the disclosure herein, for the production of identifiable polyketides, polypeptides or
polyketide/polypeptide hybrids.

Host cells (e.g. Streptomyces) can be transformed with one or more vectors,
collectively encoding a functional PKS/NRPS set, or a cocktail comprising a random

25 assortment of enediyne ORFs and/or PKS and/or NRPS genes, modules, active sites, or

portions thereof The vector(s) can include native or hybrid combinations of enediyne ORFs,
and/or PKS and/or NRPS subunits or cocktail components, or mutants thereof As explained
above, the gene cluster need not correspond to the complete native gene cluster but need only
encode the necessary enediyne ORFs and/or PKS and/or NRPS components to catalyze the

30 production of the desired product(s).

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IV. Variation of starter and/or extender units, and/or host cells.

In addition to varying the nucleic acids comprising the subject gene cluster,
variations in the products produced by the gene cluster(s) can be obtained by varying the the
host cell, the starter units and/or the extender units. Thus, for example different fatty acids
5 can be utilized in the enediyne synthetic pathway resulting in different enediyne variants.
Similarly different intermediate metabolites can be provided (e.g. endogenously produced by
the host cell, or produced by an introduced herterologous construct, and/or supplied from an
exogenous source (e.g. the culture media)). Similarly, varying the host cell can vary the
resulting product(s). For example, a gene cassette carrying the enediyne biosynthesis genes
1 0 can be introduced into a deoxysugar-synthesizing host for the production of glycosylated
enediyne metabolites.

V. Use of C-1027 resistance genes.

The antibiotic C-1027 and metabolites present in C-1027 biosynthesis are
highly potent cytotoxins. Accordingly the biosynthesis of C-1027 is facilitated by the

15 presence of one or more antibiotic (e.g. enediyne) resistance genes. Without being bound to
a particular theory, it is believed that CagA and SgcB function cooperatively to provide
resistance. It is believed that the C-1027 chromophore is first sequestered by binding to the
preaproprotein CagA (ORF 9) to form a complex, which is then transported out of the cell by
the efflux pump SgcB (ORF 2) and processed by removing the leader peptide to yield the

20 chromoprotein. Other genes that appear to mediate resistance in the C- 1 027 biosynthesis

gene cluster include a transmembrane transport protein (ORF 27), a Na + /H + transporter (ORF
0), an ABC transporter (ORF -1, C-terminus), a glycerol phosphate transporter (ORF -2), and
a UvrA-like protein (ORF -1, N-terminus) (see, e.g., Table 2).

These ORFs and/or the polypeptides encoded by these ORFs can be utilized

25 alone, or in combination with one or more other C-1027 ORFs to confer resistance to

enediyne or enediyne metabolites on a cell. This is useful in a wide variety of contexts. For
example, to increase production of enediynes. For example, it is believed that C-1027
resistance could be a limiting factor at the onset of C-1027 production. Provision of an extra
copy of the plasmid-born sgcB, and overexpression of sgcB under the control of the

30 constitutive erniE* promoter resulted in increase of C-1027 production (see example 1).

In a therapeutic context, it is sometimes desirable to confer resistance on

certain vulnerable cells. Thus, for example, where an enediyne is used as a

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chemotherapeutic, transfection of vulnerable, but healthy cells (e.g. liver cells remote from
the tumor site, stem cells, etc.) with vector(s) expressing the resistance gene(s) permits
administration of the enediyne at a higher dosage with fewer adverse effects to the organism.
Such approaches have been taken using the multi-drug resistance gene (MDR1) expressing
5 p-glycoprotein.

In another embodiment vectors are provided containing one or more
resistance genes of this invention under control of a constitutive and/or inducible promoter
thereby providing a "ready-made" expression system suitable for the expression of an
enediyne or enediyne metabolite at high concentration.
10 It is also noted that the resistance genes are expected to confer resistance to

compounds other than enediynes. The resistance genes are expected to confer resistance to
essentially any cytotoxic compound that can act as a substrate for the resistance gene(s) of
this invention.

VI. Kits.

15 In still another embodiment, this invention provides kits for practice of the

methods described herein. In one preferred embodiment, the kits comprise one or more
containers containing nucleic acids encoding one or more of the C-1027 biosynthesis gene
cluster open reading frames. Certain kits may comprise vectors encoding the sgc gene
cluster orfs and/or cells containing such vectors. The kits may optionally include any

20 reagents and/or apparatus to facilitate practice of the methods described herein. Such

reagents include, but are not limited to buffers, labels, labeled antibodies, bioreactors, cells,
etc.

In addition, the kits may include instructional materials containing directions
{i.e., protocols) for the practice of the methods of this invention. Preferred instructional

25 materials provide protocols utilizing the kit contents for creating or modifying C-1027 gene
cluster and/or for synthesizing or modifying a molecule using one or more sgc gene cluster
ORFs. While the instructional materials typically comprise written or printed materials they
are not limited to such. Any medium capable of storing such instructions and
communicating them to an end user is contemplated by this invention. Such media include,

30 but are not limited to electronic storage media (e.g., magnetic discs, tapes, cartridges, chips),
optical media (e.g., CD ROM), and the like. Such media may include addresses to internet
sites that provide such instructional materials.

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EXAMPLES

The following examples are offered to illustrate, but not to limit the claimed

invention.

Example 1

5 Genes for production of the enedivne antitumor antibiotic C-1027 in Strevtomyces
zlobisvorus are clustered with the cagA gene that encodes the C-1027 apoprotein

We have been studying the biosynthesis of C-1027 in Streptomyces
globisporus C-1027 as a model for the enediyne family of antitumor antibiotics (Thorson et
al. (1999) Bioorg. Chem., 27: 172-188). C-1027 consists of a non-peptidic chromophore and

10 an apoprotein, CagA [also called C-1027AG (Otani et al (1991) Agri Biol Chem. 55: 407-
417)]. The C-1027 chromophore is extremely unstable in the protein-free state, the structure
of which was initially deduced from an inactive but more stable degradation product
(Minami et al (1993) Tetrahedron Lett 34: 2633-2636) and subsequently confirmed by
spectroscopic analysis of the natural product (Yoshida et al (1993) Tetrahedron Lett. 34:

15 2637-2640) (Fig. 1). While the absolute stereochemistry of the deoxy sugar moiety was
established by total synthesis (Iida et al (1993) Tetrahedron Lett 34: 4079-4082), the 8S,
9S, 13S and 17 R configuration of the C-1027 chromophore were based only on computer
modeling (Okuno et al (1994) J. Med. Chem. 37: 2266-2273). Although no biosynthetic
study has been carried out specifically on C-1027, the polyketide origin of the enediyne

20 cores has been implicated by feeding experiments with 13 C-labeled acetate for the

neocarzinostatin chromophore A (Hensens et al (1989) J. Am. Chem. Soc. Ill: 3295-3299),
dynemicin (Tokiwa et al (1992) J. Am. Chem. Soc. 1 14: 4107-41 10), and esperamicin (Lam
et al (1993) Am. Chem. Soc. 115: 12340-12345); and deoxysugar biosynthesis has been
well characterized in actinomycetes (Liu and Thorson (1994) Ann. Rev. Microbiol. 48: 223-

25 256; Piepersberg (1997) pp. 81-163. In Biotechnology of antibiotics, 2nd ed. W. R. Strohl
(ed). Marcel Dekker, New York). Given the structural similarity of C-1027 to the other
enediyne cores and to deoxysugars found in other secondary metabolites, we decided to
clone either a PKS or a deoxysugar biosynthesis gene as the first step of identifying the C-
1027 gene cluster from S. globisporus.

30 Furthermore, the CagA apoprotein of C-1027 has been isolated, its amino acid

sequence has been determined, and the corresponding cagA gene has been cloned and
sequenced (Otani et al (1991) Agri. Biol Chem. 55: 407-417; Sakata et al (1992) Biosci.

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Biotech. Biochem. 56: 1592-1595). Since genes encoding secondary metabolite production
in actinomycetes have invariably been found to be clustered in one region of the microbial
chromosome (Hopwood (1997) Chem. Rev. 97: 2465-2497), we further reasoned that
mapping the cagA gene with either a putative PKS gene, a deoxysugar biosynthesis gene, or
5 both to the same region of the S. globisporus chromosome should be viewed as strong

evidence supporting the proposition that the cloned genes constitute the C-1027 biosynthesis
gene cluster.

We report here the cloning and sequencing of two genes, sgcA (Streptomyces
globisporus C-1027) and sgcB y that encode a dNDP-glucose 4,6-dehydratase (NGDH) and a

10 transmembrane efflux protein, respectively. The sgcA,B locus is indeed clustered with the
cagA gene, leading to the localization of a 75-kb gene cluster from S. globisporus. The
involvement of the cloned gene cluster in C-1027 biosynthesis was demonstrated by
disrupting the sgcA gene to generate C-1027-nonproducing mutants and by complementing
the sgcA mutants in vivo to restore C-1027 production. Our results, together with similar

15 effort in the Thorson laboratory on the calicheamicin gene cluster (Thorson et ah (1999)
Bioorg. Chem., 27: 172-188), represent the first cloning of a gene cluster for enediyne
antitumor antibiotic biosynthesis.

Materials and methods.

Bacterial strains and plasmids.

20 Escherichia coli DH5a was used as a general host for routine subcloning

(Sambrook et ah (1989) Molecular cloning, a laboratory manual. Cold Spring Harbor
Laboratory, Cold Spring Harbor, NY). E. coli XL 1-Blue MR (Stratagene, La Jolla, CA)
was used as the transduction host for cosmid library construction. E. coli SI 7-1 was used as
the donor host for E. coli-S. globisporus conjugation (Mazodier et ah (1989) J. Bacterioh

25 171: 3583-3585). Micrococcus luteus ATCC943 1 was used as the testing organism to assay
the antibacterial activity of C-1027 (Hu et ah (1988) J. Antibiot. 41: 1575-1579). The
pGEM-3zf, -5zf, and -7zf and pGEM-T vectors were from Promega (Madison, WI). S.
globisporus strains and other plasmids in this study are listed in Table 3

30 Table 3. Strains and plasmids.

Strain or Relevant Characteristics

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plasmid

S. globisporus

C-1027 Wild-type (Hu et al (1988) J. Antibiot 41: 1575-1579)

AF40 Mutant resulted from acriflavine treatment of S. globisporus C-1027, C-

1027-nonproducing (Mao et al. (1997) Chinese J. Biotechnol. 13: 195-

199)

AF44 Mutant resulted from acriflavine treatment of S. globisporus C-1027, C-

1027-nonproducing (Mao et al, supra)
AF67 Mutant resulted from acriflavine treatment of S. globisporus C-1027, C-

1027-nonproducing (Mao et al, supra)
SB1001 5gc^4-disrupted mutant resulted from integration of pBS1012 into S.

globisporus C-1027 Apr R ? C-1027-nonproducing
SB 1002 sg<^4-disrupted mutant resulted from integration of pBS1013 into S.

globisporus C-1027 Apr R , C-1027-nonproducing

Plasmids:

pOJ446 E. coli-Streptomyces shuttle cosmid, Apr R (Bierman et al. (1992) Gene, 1 16: 43-
pOJ260 E. coli vector, non-replicating in Streptomyces, Apr R (Bierman et al. supra)
pKCl 139 E. coli-Streptomyces shuttle vector, rep TS , Apr 11 (Bierman et al supra)
pWHM3 £. coli-Streptomyces shuttle vector, Th R (Vara et al ( 1 989) J. Bacteriol 111:
5872-5881)

pWHM79 erm E* promoter in pGEM-3zf (Shen and Hutchinson ( 1 996) Proc. Natl Acad.

Set USA 93: 6600-6604)
pBSlOOl 0.75-kb PCR product amplified from S. globisporus with type I PKS primers in pGEM-

T

pBS1002 0.55-kb PCR product amplified from S. globisporus with NGDH gene primers in
pGEM-T

pBS1003 0.73-kb PCR product amplified from pBS1005 with cagA primers in pGEM-T

pBS 1004 pOJ446 S. globisporus genomic library cosmid

pBS1005 pOJ446 iS. globisporus genomic library cosmid

pBS 1006 pOJ446 S. globisporus genomic library cosmid

pBS1007 3.0-kb BamHI fragment from pBS1005 in pGEM-3zf, sgcA, sgcB

pBS 1008 4.0-kb BamHI fragment from pBS 1 005 in pGEM-3zf, cagA

pBS1009 1.0-kb Kpnl truncated fragment of sgcA from pBS1007 in pGEM-3zf

pBSlOlO 0.75-kb SaclVSphl internal fragment of sgcA from pBS1009 in pGEM-5zf

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pBSlOl 1 0.75-kb SacllSphl internal fragment of sgcA from pBSlOlO in pGEM-3zf
pBS 1012 0.75-kb EcoRI/Hin dill internal fragment of sgcA from pBS 1 0 1 0 in pO J260
pBS1013 0.75-kb EcoBJ/HindUI internal fragment of sgcA from pBSlOlO in pKCl 139
pBS1014 2.0-kb EcoRI/Sphl fragment from pBS1007 in the SmaVSphl sites of pWHM79, ermE*
sgcA

pBS1015 2.5-kb EcdRUHindm fragment from pBS1014 in pWHM3, ermE* sgcA
pBS1016 Self-ligation of the 5.2-kb Kpn\ fragment from pBS1007

pBS1017 0.45-kb EcoKl/Sacl fragment from pWHM79 in EcoRUSacI sites of pBS1016, ermE*
sgcB

pBS1018 2.5-kb EcoRI/Hindlll fragment from pBS1017 in pKCl 139, ermE* sgcB

Biochemicals and chemicals.

Ampicillin, apramycin, nalidixic acid, and thiostrepton were from Sigma (St.
Louis, MO). Unless specified otherwise, restriction enzymes and other molecular biology
reagents were from standard commercial sources.

Media and culture conditions.

E. coli strains carrying plasmids were grown in Luria-Bertani (LB) medium
and were selected with appropriate antibiotics. S. globisporus strains were grown on ISP-4
(Difco Laboratories, Detroit, MI) or R2YE at 28°C for sporulation and in TSB (Hopwood et
al (1985) Genetic manipulation of Streptomyces: a laboratory manual. John Innes
Foundation, Norwich, UK) supplemented with 5 mM MgCk and 0.5% glycine at 28°C, 250
rpm for isolation of genomic DNA. For transformation, S. globisporus strains were grown in
YEME (Hopwood et al, supra.) for preparation of protoplasts and on R2YE for protoplast
regeneration. For conjugation, both the E. coli SI 7-1 donors and the S. globisporus
recipients (upon germination in TSB) were prepared in LB, and donors/recipients were
grown on either ISP-4 medium with 0.05% yeast extract and 0.1% tryptone or AS-1 medium
(Baltz (1980) Dev. Ind. Microbiol 21: 43-54; Bierman et al (1992) Gene 116: 43-69) at
30°C for isolation of exconjugants.

For C-1027 production, S. globisporus strains were grown either on R2YE or

ISP-4 agar medium at 28°C or in liquid medium by a two-stage fermentation. For liquid

culture, the seed inoculum was prepared by inoculating 50 mL medium (consisting of 2%

glycerol, 2% dextrin, 1% fish meal, 0.5% peptone, 0.2% (NH 4 ) 2 S0 45 and 0.2% CaC0 3 , pH

7.0) with an aliquot of spore suspension, incubating at 28°C, 250 rpm for two days. To a

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fresh 50 mL of the same medium was then added the seed culture (5%), and incubation
continued at 28°C, 250 rpm for three to six days (Hu et al (1988) J. Antibiot 41: 1575-
1579). The fermentation supernatants were harvested by centrifugation (Eppendorf 5415C,
4°C, 10 min, 14,000 rpm) on day 3, 4 and 5, and assayed for their antibacterial activity
5 against M. luteus (Hu et al (1988) J, Antibiot, 41: 1575-1579).

DNA isolation and manipulation.

Plasmid preparation and DNA extraction were carried out by using
commercial kits (Qiagen, Santa Clarita, CA). Total S. globisporus DNA was isolated
according to literature protocols (Hopwood et al (1985) Genetic manipulation of

10 Streptomyces: a laboratory manual John Innes Foundation, Norwich, UK; Rao et al (1987)
Methods Enzymol 153: 166-198). Restriction endonuclease digestion and ligation followed
standard methods (Sambrook et al (1989) Molecular cloning, a laboratory manual Cold
Spring Harbor Laboratory, Cold Spring Harbor, NY). For Southern analysis, digoxigenin
labeling of DNA probes, hybridization, and detection were performed according to the

15 protocols provided by the manufacturer (Boehringer Mannheim Biochemicals, Indianapolis,
IN).

DNA sequencing.

Automated DNA sequencing was carried out on an ABI Prism 377 DNA
Sequencer using the ABI Prism dye terminator cycle sequencing ready reaction kit and
20 AmpliTag DNA polymerase FS (Perkin-Elmer/ABI, Foster City, CA). Sequencing service
was provided by either the DBS Automated DNA Sequencing Facility, UC Davis, or Davis
Sequencing Inc. (Davis, CA). Data were analyzed by ABI Prism Sequencing 2.1.1 software
and the Genetics Computer Group program (Madison, WI).

Polymerase chain reaction (PCR)»

25 Primers were synthesized at the Protein Structure Laboratory, UC Davis,

PCR was carried out on a Gene Amp PCR System 2400 (Perkin-Elmer/ABI) with Tag
polymerase and buffer from Promega. A typical PCR mixture consisted of 5 ng of S.
globisporus genomic or plasmid DNA as template, 25 pmoles of each primers, 25 yM dNTP,
5% DMSO, 2 units of Taq polymerase, 1 x buffer, with or without 20% glycerol in a final

30 volume of 50 \xL. The PCR temperature program was as follows: initial denaturing at 94°C

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for 5 min, 24-36 cycles of 45 sec at 94°C, 1 min at 60°C, 2 min at 72°C, followed by
additional 7 min at 72°C.

For type II PKS, the following two pairs of degenerate primers were used —
5'-AGC TCC ATC AAG TCS ATG RTC GG-3* (forward, SEQ ID NO:103) / 5 5 -CC GGT
5 GTT SAC SGC GTA GAA CCA GGC G-3' (reverse, SEQ ID NO: 104) and 5'-GAC ACV
GCN TGY TCB TCV-3' (forward, SEQ ID NO: 105)/5'-RTG SGC RTT VGT NCC RCT-3'
(SEQ ID NO: 106) (B, C+G+T; N, A+C+G+T; R, A+G; S, C+G; V, A+C+G; Y, C+T)
(reverse) (Seow et al (1997) J. Bacteriol, 179: 7360-7368). No product was amplified
under all conditions tested. For type I PKS, the following pair of degenerate primers were

10 used— 5'-GCS TCC CGS GAC CTG GGC TTC GAC TC-3' (forward, SEQ ID NO:107) /
5'-AG SGA SGA SGA GCA GGC GGT STC SAC-3' (S, G+C) (reverse, SEQ ID NO: 108)
(Kakavas et al (1997) J. BacterioL, 179: 7515-7522). A distinctive product with the
predicted size of 0.75 kb was amplified in the presence of 20% glycerol and cloned into
pGEM-T according to the protocol provided by the manufacturer (Promega) to yield

15 pBSlOOl.

For NGDH, the following pair of degenerate primers were used — 5'-CS GGS
GSS GCS GGS TTC ATC GG-3' (forward, SEQ ID NO:109) / 5'-GG GWR CTG GYR
SGG SCC GTA GTT G-3' (R, A+G; S, C+G; W, A+T; Y, C+T) (reverse, SEQ ID NO:l 10)
(Decker, et al (1996) FEMS Lett, 141: 195-201). A distinctive product with the predicted

20 size of 0.55 kb was amplified and cloned into pGEM-T to yield pBS1002.

For cagA, the following pair of primers, flanking its coding region, were
used— 5 '-AG GTG GAG GCG CTC ACC GAG-3 ' (forward, SEQ ID NO: 1 1 1)/5 '-G GGC
GTC AGG CCG TAA GAA G-3' (reverse, SEQ ID NO: 112) (Sakata et al (1992) Bioscl
Biotechnol Biochern., 56: 159201595). A distinctive product with the predicted size of 0.73

25 kb was amplified from pBS 1005 and cloned into pGEM-T to yield pBS 1003.

Genomic library construction and screening.

S. globisporus genomic DNA was partially digested with Mbol to yield a
smear around 60 kb, as monitored by electrophoresis on a 0.3% agarose gel. This sample
was dephosphorylated upon treatment with shrimp alkaline phosphatase and ligated into the
30 E. coli-Streptomyces shuttle vector pOJ446 (Bierman et al (1992) Gene 116: 43-69) that was
prepared by digestion with Hpal, shrimp alkaline phosphatase treatment, and additional
digestion with BamUl. The resulting ligation mixture was packaged with the Gigapack II

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XL two-component packaging extract (Stratagene). The package mixture was transduced
into E. coli XL 1-Blue MR. The transduced cells were spread onto LB plates containing
apramycin (100 jag/mL) and incubated at 37°C overnight. The titer of the primary library
was approximately 6,000 colony- forming units per ^g of DNA. Restriction enzyme analysis
5 of twelve randomly selected cosmids confirmed that the average size of inserts was about 35
to 45 kb (Rao et al. (1987) Meth. EnzymoL, 153: 166-198).

To screen the genomic library, colonies from five LB plates containing
apramycin (100 jag/mL, with approximately 2,000 colonies per plate) were transferred to
nylon transfer membranes (Micro Separations, Inc., Westborough, MA) and screened by

10 colony hybridization with the PCR-amplified 0.55-kb NGDH fragment from pBS1002 as a
probe. The positive cosmid clones were re-screened by PCR with primers for NGDH and
confirmed by Southern hybridization (Sambrook et al., supra.). Further restriction enzyme
mapping and chromosomal walking of these overlapping cosmids led to the genetic
localization of the 75-kb sgc gene cluster, as represented by pBS1004, pBS1005, and

15 pBS1006 (Fig. 5A). A 3.0-kb BamRl fragment from pBS1005 that hybridized to the NGDH
probe was cloned into the same sites of pGEM-3zf to yield pBS1007. Similarly, a 4.0-kb
BamBl fragment from pBS1005 that hybridizes to the PCR-amplified 0.73-kb cagA probe
from pBS1003 was cloned into the same sites of pGEM-3zf to yield pBS1008 (Fig. 5B).

Generation of sgcA mutants by insert-directed homologous recombination in S.
20 globisporus.

A 1.0-kb Kpnl fragment from pBS1007, containing the C-terminal truncated
sgcA, was subcloned into pGEM-3zf to yield pBS1009. An internal fragment of sgcA was
moved sequentially as a 0.75-kb SaclVSphl fragment from pBS1009 into the same sites of
pGEM-5zf to yield pBSlOlO and as a 0.75-kb SacllSphl fragment from pBSlOlO into the
25 same sites of pGEM-3zf to yield pBS 101 1 . The latter plasmid was digested with EcoRI and
Hindlll, and the resulting 0.75-kb EcoKL/HindttI fragment was cloned into the same sites of
pOJ260 and pKCl 139 (Bierman et al. (1992) Gene, 116: 43-69 to yield pBS1012 and
pB S 1 0 1 3 , respectively.

Introduction of pBS1012 and pBS1013 into S. globisporus was carried out by
30 either polyethyleneglycol (PEG)-mediated protoplast transformation (Hopwood et al (1985)
Genetic manipulation of Streptomyces: a laboratory manual John Innes Foundation,
Norwich, UK) or E. coli-S. globisporus conjugation (Bierman et al. (1992) Gene 116: 43-69;

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Matsushima and Baltz (1996) Microbiology 142: 261-267; Matsushima et al (1994) Gene
146: 39-45), methods for both of which were developed recently in our laboratory. In brief,
for transformation, pBS1012 and pBS1013 were propagated in E. coli ET12567 (MacNeil et
al (1992) Gene 111: 61-68), and the resulting double strand plasmid DNA was denatured by
5 alkaline treatment (Ho and Chater (1997) J. Bacteriol 179: 122-127). The latter DNA (5
nL) and 200 jiL of 25% PEG 1000 in P buffer (Hop wood et al supra) were sequentially
added to 50 \xL of S. globisporus protoplasts (10 9 ) in P buffer. The resulting suspension was
mixed immediately and spread on R2YE plates. After incubation at 28°C for 16 to 20 hrs,
the plates were overlaid with soft R2YE (0.7% agar) containing apramycin (100 ng/mL, final

10 concentration); incubation continued until colonies appeared (in 5 to 7 days). For

conjugation, E. coli S17-l(pBS1012) or J?, coli SI 7-1 (pBS1013) was grown to an OD 600 of
0.3 to 0.4. Cells from a 20-mL culture were pelleted by centrifligation, washed in LB, and
resuspended in 2 mL of LB as the E. coli donors. S. globisporus spores (10 3 to 10 9 ) were
washed, resuspended in TSB, and incubated at 50°C for 10 min to activate germination.

15 After additional incubation at 37°C for 2 to 5 hrs, the spores were pelleted and resuspended
in LB as the S. globisporus recipients. The donors (100 jliL) and recipients (100 ^L) were
mixed and spread equally onto two modified ISP-4 or AS-1 plates supplemented freshly with
10 mM MgCl 2 (see Media and culture conditions). The plates were incubated at 28°C for 16
to 22 hrs. After removal of most of the E. coli SI 7-1 donors by washing the surface with

20 sterile water, the plates were overlaid with 3 mL of soft LB (0.7% agar) containing nalidixic
acid (50 ng/mL, final concentration) and apramycin (100 p,g/mL, final concentration) and
incubated at 28°C until exconjugants appeared (in approximately 5 days).

Unlike pBS1012, which is a Streptomyces non-replicating plasmid, pBS1013
bears a temperature-sensitive Streptomyces replication origin (Bierman et al (1992) Gene

25 116: 43-69; Muth et al (1989) Mol Gen. Genet 219: 341-348) that is unable to replicate at
temperatures above 34°C (Table 3), while the S. globisporus wild-type strain grows normally
up to 37°C. Thus, spores of S. globisporus (pBS1013), from either the transformants or the
exconjugants, were spread onto R2YE plates containing apramycin (100 ng/mL). The plates
were incubated directly at 37°C, and mutants, resulting from single crossover homologous

30 recombination between pBS1013 and the S. globisporus chromosome, were readily obtained
in 7 to 10 days. Alternatively, the plates were first incubated at 28°C for 2 days until
pinpoint-size colonies became visible and then shifted to 37°C to continue incubation.

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Mutants resulting from single crossover homologous recombination grew out of the original
pinpoint-size colonies as easily distinguishable sectors in 7 to 10 days.

Construction of the s2cA and sscB expression plasmids.

pBS1007 was digested with EcdRI, and made blunt-ended by treatment with
5 the Klenow fragment of DNA polymerase L Upon additional digestion with Sphl, the

resulting 2.0-kb blunt-ended Sphl fragment containing the intact sgcA gene was cloned into
the Small Sphl sites of pWHM79 (Shen et al (1996) Proc. Natl Acad. Set, USA, 93: 6600-
6604) to yield pBS1014. The latter was digested with EcoBl and Hindlll, and the resulting
2.5-kb EcoRIIHindlll fragment was cloned into the same sites of pWHM3 (Vara et al

10 (1989) J. Bacteriol 171: 5872-5881) to yield pBS1015, in which the expression of sgcA is
under the control of the ermE* promoter (Bibb et al (1994) Mol Microbiol 14: 533-545).

Alternatively, pBS1007 was digested with Kpnl, removing most of the sgcA
gene, and the 5.2-kb Kpnl fragment was recovered and self-ligated to yield pBS1016. The
ermE* promoter was subcloned from pWHM79 (Shen et al (1996) Proc. Natl Acad. Set,

15 USA, 93: 6600-6604) as a 0.45-kb EcoKUSacl fragment and cloned into the same sites of
pBS1016 to yield pBS1017. The latter was digested with EcoRI and Hindlll, and the
resulting 2.5-kb EcoSllHindlll fragment was cloned into the same sites of pKCl 139 to yield
pBS1018, in which the expression of sgcB is under the control of the ermE* promoter.

Determination of C-1027 production.

20 The production of C-1027 was detected by assaying its antibacterial activity

against M. luteus (Hu et al (1988) J. Antibiot. 41: 1575-1579). From liquid culture,
fermentation supernant (180 ^iL) was added to stainless steel cylinders placed on LB plates
pre-seeded with overnight M. luteus culture (0.01% vol/vol). From solid culture, a small
square block (0.5 x 0.5 x 0.5 cm 3 ) of agar from either R2YE or ISP-4 medium was directly

25 placed on M. luteus-seeded LB plates. The plates were incubated at 37°C for 24 hrs, and C-
1027 production was estimated by measuring the size of inhibition zones.

Nucleotide sequence accession number.

The nucleotide sequence reported here has been deposited in the GenBank
database with the accession number AF201913.

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Results.

No polyketide synthase gene was amplified by PCR from S. slobisvorus.

On the assumption that the C-1027 enediyne core is of polyketide origin, the
PCR approach was adopted to screen S. globisporus for any putative PKS genes, although it
5 is far from certain a priori if the biosynthesis of the enediyne core invokes a PKS and, if so,
whether the enediyne PKS will exhibit a type I or type II structural organization. PCR
methods for cloning either type I or type II PKS genes have been developed, and these
methods have proven to be very effective in cloning PKS genes from various polyketide-
producing actinomycetes (Kakavas et ah (1997) J. BacterioL 179: 7515-7522; Seow et ah

10 (1997) J. Bacterioh 179: 7360-7368). While no distinctive product was amplified under all
conditions examined with both pairs of primers designed for type II PKS, a single product
with the expected size of 0.75 kb was readily amplified by PCR from S. globisporus with
primers designed for type I PKS, which was subsequently cloned (pBSlOOl). Intriguingly,
sequence analysis of six randomly selected pBSlOOl clones yielded an identical product —

15 indicative of a specific PCR amplification — the deduced amino acid sequence of which,
however, showed no homology to known PKSs (data not shown), excluding the possibility
of using PKS as a probe to identify the sgc biosynthesis gene cluster.

Cloning of a putative NGDH gene by PCR from S. globisporus.

The biosynthesis of various deoxyhexoses share a common key
20 intermediate — 4-keto-6-deoxyglucose nucleoside diphosphate or its analogs — whose
formation from glucose nucleoside diphosphate is catalyzed by the NGDH enzyme, an
NAD + -dependent oxidoreductase (Liu and Thorson (1994) Ann. Rev. Microbiol. 48: 223-
256; Piepersberg (1997) pp. 81-163. In Biotechnology of antibiotics, 2nd ed. W. R. Strohl
(ed). Marcel Dekker, New York). The PCR method was adopted to clone the putative
25 NGDH gene from S. globisporus with primers designed according to the homologous

regions of various NGDH enzymes from actinomycetes (Decker et ah (1996) FEMSLett.
141: 195-201), resulting in the amplification of a single product with the expected size of
0.55 kb (pBS1002). Sequence analysis of pBS1002 confirmed its identity as a part of a
putative NGDH gene.

30 To clone the complete NGDH gene, an S. globisporus genomic library,

constructed in the E. coli-Streptomyces shuttle vector pOJ446 (Bierman et ah (1992) Gene

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116: 43-69; Rao et al (1987) Methods Enzymol 153: 166-198), was analyzed by Southern
hybridization with the PCR-amplified 0.55-kb fragment from pBS1002 as a probe. Of the
10,000 colonies screened, 36 positive colonies were identified, 9 of which were confirmed
by PCR to harbor the DGDH gene. Restriction enzyme mapping showed that all of them
5 contained a single 3.0-kb BamUl fragment hybridizing to the NGDH probe. Additional

chromosomal walking from this locus eventually led to the localization of the 75-kb sgc gene
cluster, covered by 18 overlapping cosmids as represented by pBS1004, pBS1005, and
pBS1006 (Fig. 5 A). The 3.0-kb BamHl fragment was subcloned (pBS1007) (Fig. 5B), and
its nucleotide (nt) sequence was determined.

10 Analysis of the DNA sequences of the sgcA and sscB genes.

Two complete open reading frames (ORFs) (sgcA and sgcB) were identified
within the 3.0-kb BamHl fragment of pBS1007, the 3,035-nt sequence of which is shown in
Figure 6. The sgcA gene most likely begins with an ATG at nt 101, preceded by a probable
ribosome biding site (RBS), GGAGG, and ends with a TGA stop codon at nt 1099. SgcA

15 should therefore encode a 332-amino acid protein with a molecular weight of 36,341 and an
isoelectric point of 6.01. A Gapped-BLAST search showed that the deduced sgcA gene
product is highly homologous to various putative and known NGDH enzymes from
antibiotic-producing actinomycetes, including Gdh from the erythromycin biosynthesis gene
cluster in Saccharopolyspora erythraea (64% identity and 70% similarity) (Linton et al

20 (1995) Gene 153: 33-40), MtmE from the mithramycin biosynthesis gene cluster in
Streptomyces argillaceus (64% identity and 68% similarity) (Lombo et al (1997) J.
BacterioL 179: 3354-3357), and TylA2 from the tylosin biosynthesis gene cluster in
Streptomyces fradiae (62% identity and 68% similarity) (Merson-Davies and Cundliffe
(1994) Mot Microbiol 13: 349-355) (Fig. 7). A conserved sequence of 14 amino acid

25 residues close to the N-termini can be easily identified in these proteins, which has been
described as a pap fold with an NAD + -binding motif, GxGxxG, (Fig. 7, boxed), consistent
with their biochemical role in deoxyhexose biosynthesis (Liu and Thorson (1994) Ann. Rev.
Microbiol 48: 223-256; Piepersberg (1997) pp. 81-163. In Biotechnology of antibiotics, 2nd
ed. W. R. Strohl (ed). Marcel Dekker, New York). The function of Gdh and MtmE as TDP-

30 glucose 4,6-dehydratases, requiring NAD + as a cofactor, has been confirmed by an enzyme
assay following expression of the gdh (Linton et al (1995) Gene 153: 33-40) and mtmE gene
(Lombo et al (1997) J. Bacteriol 179: 3354-3357) in E. coli, respectively, and by

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purification of the Gdh protein from Sacc. erythraea (Vara et al (1989) J. Bacteriol 171:
5872-5881). From these data, it is reasonable to suggest that sgcA encodes the NGDH
enzyme required for the biosynthesis of the 4,6-dideoxy-4-dimethylamino-5-
methylrhamnose moiety of the C-1027 chromophore.
5 Transcribed in the same direction as sgcA, the sgcB gene is located 43 nt

downstream of sgcA. It should begin with a GTG at nt 1 143, preceded by a probable RBS,
AGGAG, and end with a TGA at nt 2708 (Fig. 6). Correspondingly, sgcB should therefore
encode a 521 -amino acid protein with a molecular weight of 52,952 and an isoelectric point
of 4.64. Database comparison of the deduced sgcB product revealed that SgcB is closely
10 related to a family of membrane efflux pumps, such as LfiA from Mycobacterium smegmatis
(43% identity and 50% similarity, protein accession number AAC43550) (Takiff et al

(1996) Proc. Natl Acad. Set USA 93: 362-366), OrfA from Streptomyces cinnamomeus
(42% identity and 47% similarity, protein accession number AAB71209) (Sommer et al

(1997) Appl Environ. Microbiol 63: 3553-3560), and RifiP from the rifamycin biosynthesis
1 5 gene cluster in Amycolatopsis mediterranei (35% identity and 44% similarity, protein

accession number AAC01725) Augus et al (1998) Chem. Biol 5: 69-79). These proteins are
membrane-localized transporters involved in the transport of antibiotics (conferring
resistance), sugars, and other substances. While direct evidence is lacking for RifP
conferring rifamycin resistance in A. mediterranei by transporting it out of the cells (August

20 et al (1998) Chem. Biol, 5: 68-79), it has been proven that LfrA employs the

transmembrane proton gradient in an antiporter mode to drive the efflux of intracellular
antibiotics, resulting in fluoroquinolone resistance inM smegmatis (Takiff et al (1996)
Proc. Natl Acad. Scl USA 93: 362-366). On the basis of the high degree of amino acid
sequence conservation, an equivalent role could be proposed for SgcB, conferring resistance

25 by exporting C-1027 from S. globisporus.

The cagA gene is clustered with the S2CA and sscB locus.

To determine if cagA is clustered with the sgcA and sgcB locus, PCR primers
were designed according to the flanking regions of cagA (Sakata et al (1992) Bioscl
Biotech. Biochem. 56: 1592-1595). A single product with the predicted size of 0.73 kb was
30 indeed amplified from several of the overlapping cosmids (which cover the 75-kb sgc

cluster), including pBS1004 and pBS1005, the identity of which as cagA was confirmed by
sequencing. Restriction enzyme mapping and Southern hybridization analysis localized

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cagA to a single 4.0-kb BamRl fragment that is approximately 14 kb upstream of the sgcA,B
locus (Fig. 5B). The 4.0-kb BamRl fragment was subcloned (pBS1008), and its nt sequence
was determined, revealing the cagA gene along with two additional ORFs (data not shown)
(Fig. 5). As reported earlier, cagA encodes a 142-amino acid protein that is processed by
5 cleavage of a 32-amino acid lead peptide to yield the mature CagA apoprotein (Sakata et al
(1992) BioscL Biotech. Biochem. 56: 1592-1595).

Disruption of the sgcA gene in S. globisporus.

To examine if the cloned sgc cluster encodes C-1027 biosynthesis, sgcA was
insertionally disrupted by a single crossover homologous recombination event to generate C-

10 1027-nonproducing mutant strains (Fig. 8 A). Two plasmids were used — pBS1012 (a

pOJ260 derivative) and pBS1013 (a pKCl 139 derivative), each of which contain a 0.75-kb
internal fragment from sgcA (Table 3). After introduction of pBS1012 into S. globisporus
either by PEG-mediated protoplast transformation or E. coli-S. globisporus conjugation,
transformants or exconjugants that were resistant to apramycin were isolated in all cases.

15 Since pBS1012 is derived from the Streptomyces non-replicating plasmid of pOJ260, these
isolates must have resulted from integration of pBS1012 into the S. globisporus chromosome
by homologous recombination. Plasmid pBS1013 was similarly introduced into S.
globisporus. However, since pBS1013 is derived from pKCl 139 that carries the
temperature-sensitive Streptomyces replication origin from pSG5 and can replicate normally

20 at 28°C (Muth et al (1989) Mol Gen. Genet 219: 341-348), these isolates were subjected to
incubation at the non-permissive temperature of 37°C to eliminate free plasmids from the
host cells. As expected, normal growth stopped except for the recombinants that continue to
grow at 37°C 5 indicative of integration of pBS1013 into S. globisporus by homologous
recombination. The apramycin-resistant S. globisporus SB 1001 and S. globisporus SB 1002

25 strains were chosen as representatives of mutant strains with disrupted sgcA gene resulted
from integration of pBS1012 and pBS1013, respectively.

To confirm that targeted sgcA disruption has occurred by a single crossover
homologous recombination event, Southern analysis of the DNA from the mutant strains was
performed as exemplified for S. globisporus SB 1001 with either pOJ260 or the 0.75-kb

30 Sacll/Kpnl internal fragment of sgcA from pBSlOlO as a probe. As shown in Fig. 8B, a
distinctive band of the predicted size of 6.3 kb was detected with the pOJ260 vector as a
probe in all mutant strains (lanes 2, 3, and 4); this band was absent from the wild-type strain

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(lane 1). Complementarity, when using the 0.75-kb SaclllKpnl internal fragment of sgcA as
a probe (Fig. 8C), the 3.0-kb band in the wild-type strain (lane 1) was split into two
fragments with the size of 6.3 kb and 1 .0 kb in the mutant strains (lanes 2, 3, and 4), as
would be expected for disruption of sgcA by a single crossover homologous recombination
event.

S. globisporus SB1001 and S. zlobisvorus SB1002 are C-1027-nonproducing
mutants.

No apparent difference in growth characteristics and morphologies between
the wild-type S. globisporus and mutant S. globisporus SB 1001 and S. globisporus SB 1002
strains was observed. While C-1027 production in the wild-type S. globisporus strain could
be detected on day 3, peaked on day 5, and continued for a few more days, as judged by
assaying the antibacterial activity of the culture supernant against M luteus (Hu et al (1988)
J. AntibioL 41: 1575-1579), C-1027 production is completely abolished in the sgcA mutant
strains S. globisporus SB 1001 and S. globisporus SB 1002 (Fig. 9A). The latter phenotype
was identical to that of the AF40, AF44, and AF67 mutants, C-1027-nonproducing S.
globisporus strains that have been characterized previously (Fig. 9A and 9C) (Mao, et al
(1997) Chinese J. BiotechnoL 13: 195-199).

In vivo complementation of S. globisporus SB1001.

The ability of the wild-type sgcA gene to complement the disrupted sgcA gene
was tested in the S. globisporus SB1001 strain. The construction of pBS1015, in which the
expression of sgcA is under the control of the constitutive errnE* promoter, was described in
Materials and Methods. Both the pBS1015 construct and the pWHM3 vector as a control
were introduced by transformation into the S. globisporus SB 1001 mutant strains. Culture
supernants from each transformant were bioassayed against M. luteus for C-1027 production.
pBS1015 restored C-1027 production to 5. globisporus SB 1001 to the wild-type level; no C-
1027 production was detected in the control in which pWHM3 was introduced into S.
globisporus BS1001 (Fig. 9B and 9C). A significant reduction of C-1027 production was
observed when S. globisporus SB1001(pBS1015) was cultured under identical conditions but
without thiostrepton (Fig. 9B vs. 6C), indicative that pBS1015 may be unstable in S.
globisporus SB 1001 in the absence of antibiotic selection pressure.

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Expression of sgcB in S. globisporus.

The effect of sgcB on C-1027 production was tested in the wild-type S.
globisporus strain. The construction of pBS1018, in which the expression of sgcB is under
the control of the constitutive ermE* promoter, was described in Materials and Methods.
5 pBS1018 and the pKCl 139 vector as a control were each introduced by conjugation into S.
globisporus. Culture supernatants from each exconjugant were harvested on days 3, 4, and
5, and assayed for C-1027 production by determining the antibacterial activity against M
luteus. While no apparent difference for C-1027 production was observed between the S.
globisporus and S. globisporus (pKCl 139) strains, a significant increase in C-1027
1 0 production ( 1 50±25%) was evident in the early stage of S. globisporus (pBS 1018)

fermentation (Fig. 9D, day 3). However, such effect on C-1027 production leveled off as the
fermentation proceeded and became insignificant when the culture reached the late stationary
phase of fermentation (Fig. 9D, day 4 and 5).

Discussion.

15 Our inability to clone the putative enediyne PKS gene by PCR, with

degenerate primers designed according to the highly conserved amino acid sequences of
either type I or type II PKSs, or by DNA hybridization, with homologous type I or type II
PKS as probes (data not shown), was unexpected, since feeding experiments by
incorporation of [1- 13 C]- and [ 1, 2- 13 C] acetate into the enediyne cores of esperamicin (Lam

20 et al (1993) J. Am. Chem. Soc. 115: 12340-12345), dynemicin (Tokiwa et al. (1992) J. Am.
Chem. Soc. 1 14: 4107-41 10), and neocarzinostatin (Hensens et al (1989) /. Am. Chem. Soc.
Ill: 3295-3299) supported their polyketide origin. Although the enediyne cores are
structurally distinct from either the reduced or aromatic polyketides, the biosynthesis of
which is well characterized by type I or type II PKS, respectively, it could be imagined that

25 an enediyne PKS catalyzes the biosynthesis of a polyunsaturated linear heptaketide

intermediate that is subsequently cyclized into the enediyne core structure (Hu et al. (1994)
Mol Microbiol 14: 163-172; Spainke/a/. (1991) Nature 354: 125-130; Thorson etf
(1999) Bioorg. Chem., 27: 172-188). Alternatively, Hensens and co-workers proposed a
fatty acid origin for the enediyne core that was also consistent with the isotope labeling

30 results. These authors suggested oleate as a precursor that is shortened by loss of carbons

from both ends and is desaturated via the oleate-crepenynate pathway to furnish the enediyne

core (Hensens et al. (1989) /. Am. Chem. Soc. Ill: 3295-3299). The latter pathway

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resembles polyacetylene biosynthesis in higher plants and fungi and requires an acetylene
forming enzyme — a plant gene encoding such an enzyme was identified recently (Lee et al
(1998) Science 280: 915-918). Our DNA sequence analysis of approximately 60 kb of the
sgc gene cluster, fails to reveal any gene that resembles PKS.

Although little is known about the resistance mechanism for the enediyne
antibiotics in general, the apoproteins of the chromoprotein type of enediynes could be
viewed as resistance elements that confer self-resistance to the producing organisms by drug
sequestration (Thorson et al (1999) Bioorg. Chern., 27: 172-188). Such a resistance
mechanism is in fact well established in antibiotic-producing actinomycetes, for example,
BlmA, the bleomycin-binding protein from Streptomyces verticillus (Shen et al (1999)
Bioorg. Chem. 27: 155-171). Given the fact that antibiotic production genes have invariably
been found to be clustered in one region of the microbial chromosome, consisting of
structural, resistance, and regulatory genes, we adopted a strategy to clone the sgc gene
cluster by mapping a putative C-1027 structural gene to the previously cloned cagA gene,
considered as a resistance gene that encodes the C-1027 apoprotein.

We chose NGDH as the putative C-1027 structural gene on the basis of the
4,6-dideoxy-4-dimethylamino-5-methylrhamnose moiety of the C-1027 chromophore. It has
been well established that all deoxyhexoses could be derived from the common intermediate
of 4-keto-6-deoxy glucose nucleoside diphosphate, the biosynthesis of which from glucose
nucleoside diphosphate is catalyzed by an NGDH enzyme. We cloned the NGDH gene from
S. globisporus by PCR and used it as a probe to screen an S. globisporus genomic library,
resulting in the isolation of the 75-kb sgc gene cluster. DNA sequence analysis of a 3.0-kb
BarriHl fragment of the sgc cluster confirmed the presence of the NGDH protein, encoded by
sgcA, along with sgcB that encodes a transmembrane efflux protein (Fig. 6). The cagA gene
indeed resides approximately 14 kb upstream of sgcA (Fig. 5); DNA sequence analysis of a
4.0-kb BamBl fragment confirmed the identity of cagA along with two additional ORFs
(data not shown). These results underline once again the effectiveness of cloning natural
product biosynthesis gene clusters by exploiting the clustering phenomenon between
resistance and structural genes.

The involvement of the cloned gene cluster in C-1027 biosynthesis was

demonstrated by disrupting the sgcA gene to generate S. globisporus mutants, the ability of

which to produce C-1027 was completely abolished (Fig. 9 A), and by complementing the

sgcA mutants in vivo upon expression of sgcA in trans to restore C-1027 production (Fig. 9B

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and 6C). These data unambiguously establish that sgcA is essential for C-1027 production,

and thus support the conclusion that the cloned gene cluster encodes C-1027 biosynthesis. It

should be pointed out that, although the sgcA mutants S. globisporus SB 1001 and S.

globisporus SB 1002 were characterized as C-1027-nonproducing on the basis of the

antibacterial assay alone (Fig. 9 A), this phenotype was identical to that of the controls of the

AF40, AF44, and AF67 mutants (Fig. 9A and 9C). The latter strains were isolated

previously upon randomly mutagenizing the wild-type S. globisporus strain with acriflavine

and confirmed to be C-1027-nonproducing by both the antibacterial bioassay and an

antitumor spermatogonial assay (Mao, et al (1997) Chinese 1 Biotechnol 13: 195-199),

providing strong support to the current study. Gene disruption and complementation in S.

globisporus were made possible by the recently developed genetic system that allowed us to

introduce plasmid DNA into S. globisporus via either PEG-mediated protoplast

transformation (Hopwood et al. (1985) Genetic manipulation ofStreptomyces: a laboratory

manual John Innes Foundation, Norwich, UK) or E. coli-S. globisporus conjugation

(Bierman et al (1992) Gene 116: 43-69; Matsushima and Baltz (1996) Microbiology 142:

261-267; Matsushima et al (1994) Gene 146: 39-45) for analyzing the sgc biosynthesis gene

cluster in vivo. Given the difficulties encountered with calicheamicin biosynthesis in

Micromonospora echinospora, into which all attempts to introduce plasmid DNA have failed

(Thorson et al (1999) Bioorg. Chern., 27: 172-188), the latter results underscore the

importance of selecting C-1027 as a model system for enediyne biosynthesis so that many of

the genetic tools developed in Streptomyces species can now be directly applied to the study

of enediyne biosynthesis.

Finally, the function of sgcB was probed by examining C-1027 production,

following expression of the gene in the wild-type S. globisporus strain. Database

comparison of the deduced amino acid sequence clearly suggested SgcB as a transmembrane

efflux protein, conferring resistance by exporting C-1027 out of the cell. Hence, in addition

to CagA, SgcB could be viewed as the second resistance element identified for C-1027

biosynthesis. Multiple resistance genes have been identified in numerous antibiotic

biosynthesis gene clusters (Hopwood (1997) Chem. Rev. 97: 2465-2497). It could be

imagined that CagA and SgcB function cooperatively to provide resistance — the C-1027

chromophore is first sequestered by binding to the preaproprotein CagA to form a complex,

which is then transported out of the cell by the efflux pump SgcB and processed by removing

the leader peptide to yield the chromoprotein, although we do not have any experimental

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data to substantiate this speculation. Since it is known that yields for antibiotic production
could be profoundly altered by the introduction of extra copies of regulatory, resistance, or
structural genes into wild-type organisms (Hutchinson (1994) Bio/Technology 12: 375-380),
we tested the effect of overexpressing sgcB in S. globisporus on C-1027 production. While
5 no apparent adverse effect on C-1027 production was observed upon introduction of the
pKCl 139 vector into S. globisporus (data not shown), a significant increase in C-1027
production (150±25%) was observed in the early stage of S. globisporus (pBS1017)
fermentation (Fig. 9D, day 3), supporting the predicted function for SgcB in C-1027
biosynthesis. We propose that C-1027 resistance could be a limiting factor at the onset of C-

10 1027 production, which is circumvented by the extra copy of the plasmid-born sgcB, and
overexpression of sgcB under the control of the constitutive ermE* promoter results in
increase of C-1027 production. However, as the S. globisporus (pBS1017) fermentation
proceeds to its stationary phase, C-1027 resistance is no longer a limiting factor for overall
C-1027 production, and the effect of extra copy of SgcB on C-1027 production consequently

15 became insignificant (Fig. 9D, day 5).

In conclusion, genetic analysis of enediyne biosynthesis has heretofore met
with little success in spite of considerable effort (Thorson et al (1999) Bioorg. Chem,, 27:
172-188). The localization of the sgc gene cluster and characterization of the sgcA and sgcB
genes have now provided an excellent basis for genetic and biochemical investigations

20 and/or modification of C-1027 biosynthesis, and gene disruption and overexpression in S.
globisporus clearly demonstrated the potential to construct enediyne-overproducing strains
and to produce novel enediynes that may have enhanced potency as novel anticancer drugs
using combinatorial biosynthesis and targeted mutagenesis. We envisage that the results
from C-1027 biosynthesis should facilitate the cloning and characterization of biosynthesis

25 gene clusters of other enediyne antibiotics in Streptomyces as well as in other actinomycetes,
and could have a great impact on the overall field of combinatorial biosynthesis.

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It is understood that the examples and embodiments described herein are for
illustrative purposes only and that various modifications or changes in light thereof will be
suggested to persons skilled in the art and are to be included within the spirit and purview of
5 this application and scope of the appended claims. All publications, patents, and patent
applications cited herein are hereby incorporated by reference in their entirety for all
purposes.

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CLAIMS

What is claimed is:

1 . An isolated nucleic acid comprising a nucleic acid selected from the
group consisting of

5 a nucleic acid encoding any of C-1027 open reading frames (ORFs) -7

through 42, excluding ORF 9 (cagA);

a nucleic acid encoding a polypeptide encoded by any of C-1027 open
reading frames (ORFs) -7 through 42, excluding ORF 9 (cagA); and

a nucleic acid amplified by polymerase chain reaction (PCR) using
10 primer pairs that amplify any of C-1027 open reading frames (ORFs) -7 through 42,
excluding ORF 9 (cagA).

2. The isolated nucleic acid of claim 1, wherein said nucleic comprises a
nucleic acid encoding at least two open reading frames (ORFs) selected from the group
consisting of ORF-1 through ORF 42, excluding ORF 9 (cagA).

15 3. The isolated nucleic acid of claim 1, wherein said nucleic comprises a

nucleic acid encoding at least three open reading frames (ORFs) selected from the group
consisting of ORF-1 through ORF 42, excluding ORF 9 (cagA).

4. An isolated nucleic acid comprising a nucleic acid that specifically
hybridizes under stringent conditions to an open reading frame (ORF) of the C-1027

20 biosynthesis gene cluster, excluding ORF 9 (cagA), and can substitute for the ORF to which
it specifically hybridizes to direct the synthesis of an enediyne.

5. The isolated nucleic acid of claim 4, wherein said isolated nucleic acid
comprises a nucleic acid that specifically hybridizes under stringent conditions to a nucleic
acid selected from the group consisting of ORF -7, ORF -6, ORF -5, ORF -4, ORF -3, ORF -

25 2, ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 8, ORF 10,
ORF 11, ORF 12, ORF 13, and ORF 14.

6. The isolated nucleic acid of claim 4, wherein said isolated nucleic acid
comprises a nucleic acid that specifically hybridizes under stringent conditions to a nucleic

-55-

acid selected from the group consisting of ORF 15, ORF 16, ORF 17, ORF 18, ORF 19,
ORF 20, ORF 21, ORF 22, ORF 23, ORF 24, ORF 25, ORF 26, ORF 27, ORF 28, ORF 29,
ORF 30, ORF 31, ORF 32, ORF 33, ORF 34, ORF 35, ORF 36, ORF 37, ORF 38, ORF 39,
ORF 40, ORF 41, and ORF 42.

7. The isolated nucleic acid of claim 5, wherein said isolated nucleic acid
comprises a nucleic acid selected from the group consisting of ORF -7, ORF -6, ORF -5,
ORF -4, ORF -3, ORF -2, ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4, ORF 5, ORF 6,
ORF 7, ORF 8, ORF 10, ORF 11, ORF 12, ORF 13, and ORF 14.

8. The isolated nucleic acid of claim 6, wherein said isolated nucleic acid
comprises a nucleic acid selected from the group consisting of ORF 15, ORF 16, ORF 17,
ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23, ORF 24, ORF 25, ORF 26, ORF 27,
ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33, ORF 34, ORF 35, ORF 36, ORF 37,
ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42.

9. The isolated nucleic acid of claim 4, wherein said nucleic acid
comprises a nucleic acid that is a single nucleotide polymorphism (SNP) of a nucleic acid
selected from the group consisting of ORF -7, ORF -6, ORF -5, ORF -4, ORF -3, ORF -2,
ORF -1, ORF 0, ORF 1, ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 81, ORF 1,
ORF 2, ORF 3, ORF 4, ORF 5, ORF 6, ORF 7, ORF 8, ORF 10, ORF 11, ORF 12, ORF 13,
ORF 14, ORF 15, ORF 16, ORF 17, ORF 18, ORF 19, ORF 20, ORF 21, ORF 22, ORF 23,
ORF 24, ORF 25, ORF 26, ORF 27, ORF 28, ORF 29, ORF 30, ORF 31, ORF 32, ORF 33,
ORF 34, ORF 35, ORF 36, ORF 37, ORF 38, ORF 39, ORF 40, ORF 41, and ORF 42.

10. An isolated gene cluster comprising open reading frames encoding
polypeptides sufficient to direct the assembly of a C-1027 enediyne or a C-1027 enediyne
analogue.

1 1 . The gene cluster of claim 10, wherein said gene cluster is present in a

bacterium,

12. The gene cluster of claim 11, wherein said gene cluster is present in a
bacterium selected from the group consisting of Actinomycetes, Actinoplanetes,
Actinornadura, Micromonospora, and Streptomycetes.

-56-

13. The gene cluster of claim 11, wherein said gene cluster is present in a
bacterium selected from the group consisting Streptomyces globisporus, Streptomyces
lividans, Streptomyces coelicolor, Micromonospora echinospora spp. calichenisis,
Actinomadura verrucosopora, Micromonospora chersina, Streptomyces carzinostaticus, and

5 Actinomycete L585-6.

14. The gene cluster of claim 13, wherein one or more open reading
frames is operatively linked to a heterologous promoter.

15. An isolated polypeptide comprising a catalytic domain encoded by a
nucleic acid of a C-1027 gene cluster wherein said nucleic acid comprises a nucleic acid

10 selected from the group consisting of

a nucleic acid encoding any of C-1027 open reading frames (ORFs) -7
through 42, excluding ORF 9 (cagA); and

a nucleic acid amplified by polymerase chain reaction (PCR) using
any one of the primer pairs identified in Tables I and II that specifically amplify one or more
1 5 of (ORFs) -7 through 42, excluding ORF 9 (cagA).

16. The polypeptide of claim 15, wherein said polypeptide is encoded by
at least two open reading frames selected from the group consisting of C-1027 open reading
frames (ORFs) -7 through 42, excluding ORF 9 (cagA).

17. The polypeptide of claim 15, wherein said polypeptide is encoded by
20 at least three open reading frames selected from the group consisting of C-1027 open reading

frames (ORFs) -7 through 42, excluding ORF 9 (cagA).

18. An expression vector comprising a nucleic acid of any one of claims 1

through 9.

19. A host cell transformed with an expression vector of claim 18.

25 20. The host cell of claim 1 9, wherein said cell is transformed with an

exogenous nucleic acid comprising a gene cluster encoding polypeptides sufficient to direct
the assembly of a C-1027 enediyne or a C-1027 enediyne analogue.

21. The host cell of claim 19, wherein said host cell is a bacterium.

-57-

22. The host cell of claim 21, wherein said bacterium is selected from the
group consisting of Actinomycetes, Actinoplanetes, Actinomadura, Micromonospora, and
Streptomycetes.

23. The host cell of claim 21, wherein said bacterium is selected from the
5 group consisting of Streptomyces globisporus, Streptomyces lividans, Streptomyces

coelicolor, Micromonospora echinospora spp. calichenisis, Actinomadura verrucosopora,
Micromonospora chersina, Streptomyces carzinostaticus, and Actinomycete L585-6.

comprising contacting a biological molecule that is a substrate for a polypeptide encoded by
10 a C-1027 biosynthesis gene cluster open reading frame, with a polypeptide encoded by a C-
1 027 biosynthesis gene cluster open reading frame whereby said polypeptide chemically
modifies said biological molecule.

selected from the group consisting of a hydroxylase, a homocysteine synthase, a dNDP-
15 glucose dehydrogenase, a citrate carrier protein, a C-methyl transferase, an N-methyl
transferase, an aminotransferase, a CagA apoprotein, an NDP-glucose synthase, an
epimerase, an acyl transferase, a coenzyme F390 synthase, and epoxidase hydrolase, an
anthranilate synthase, a glycosyl transferase, a monooxygenase, a type II condensation
protein, an aminomutase, a type II adenylation protein, an O-methyl transferase, a P-450
20 hydroxylase, an oxidoreductase, and a proline oxidase.

26. The method of claim 24, wherein said method comprising contacting
said biological molecule with at least two different polypeptides encoded by C-1027
biosynthesis gene cluster open reading frames.

27. The method of claim 24, wherein said method comprising contacting
25 said biological molecule with at least three different polypeptides encoded by C-1027

biosynthesis gene cluster open reading frames.

24. A method of chemically modifying a biological molecule, said method

25. The method of claim 24, wherein said polypeptide is an enzyme

28. The method of claim 24, wherein said contacting is in a host cell.

29.

The method of claim 28, wherein said host cell is a bacterium.

-58-

30. The method of claim 24, wherein said contacting ex vivo.

3 1 . The method of claim 28, wherein said biological molecule is an
endogenous metabolite produced by said host cell

32. The method of claim 28, wherein said biological molecule is an
5 exogenous supplied metabolite.

33. The method of claim 28, wherein said host cell is a eukaryotic cell.

34. The method of claim 33, wherein said eukaryotic cell is selected from
the group consisting of a mammalian cell, a yeast cell, a plant cell, a fungal cell, and an
insect cell.

10 35. The method of claim 28, wherein said host cell synthesizes sugars and

glycosylates the biological molecule.

36. The method of claim 35, wherein said host cell synthesizes

deoxy sugars.

37. The method of claim 24, wherein said method further comprises
15 contacting said biological molecule with a polyketide synthase or a non-ribosomal

polypeptide synthetase.

38. The method of claim of claim 24, wherein said contacting is in a

bacterial cell.

39. The method of claim of claim 24, wherein said contacting is ex vivo,

20 40. The method of claim 24, wherein said method comprises contacting

said biological molecule with at substantially all of the polypeptides encoded by C-1027
biosynthesis gene cluster open reading frames and said method produces an enediyne or
enediyne analogue.

41. The method of claim 24, wherein said biological molecule is a fatty
25 acid and said biological molecule is contacted with a C-1027 orf polyeptide selected from the

-59-

group consisting of an epoxide hydrase, a monooxygenase, an iron-sulfer flavoprotein, a p-
450 hydroxylase, an oxidoreductase, and a proline oxidase.

42. The method of claim 41, wherein said biological molecule is a fatty
acid and said biological molecule is contacted with a plurality of C-1027 orf polypeptides
comprising an epoxide hydrase, a monooxygenase, an iron-sulfer flavoprotein, a p-450
hydroxylase, an oxidoreductase, and a proline oxidase.

43. The method of claim 42, wherein said biological molecule is contacted
with polypeptides encoded by ORF17, ORF20, ORF21, ORF29, ORF30, ORF32, ORF35,
and ORF38.

44. The method of claim 41, wherein said biological molecule is contacted
with polypeptides encoded by ORF 15, ORF 16, ORF 28, ORF3, ORF 14, and ORF 13.

45. The method of claim 44 wherein said biological molecule is also
contacted with polypeptides encoded by ORF 4 and ORF 3.

46. The method of claim 24, wherein said method comprises contacting a
sugar with one or more C-1027 open reading frame polypeptides selected from the group
consisting of a dNDP-glucose synthase, a dNDP glucose dehydratase, an epimerase, an
aminotransferase, a C-methyltransferase, an N-methyltransferase, and a glycosyl transferase.

47. The method of claim 46, wherein said method comprises contacting a
dNDP-glucose with a plurality of C-1027 open reading frame polypeptides comprising a
dNDP-glucose synthase, a dNDP glucose dehydratase, an epimerase, an aminotransferase, a
C-methyltransferase, an N-methyltransferase, and a glycosyl transferase.

48. The method of claim 24, wherein said method comprises contacting an
amino acid with one or one or more C-1027 open reading frame polypeptides selected from
the group consisting of a hydroxylase, an aminomutase, a type II NRPS condensation
enzyme, a type II NRPS adenylation enzyme, and a type II peptidyl carrier protein.

49. The method of claim 48, wherein said method comprises contacting an
amino acid with a plurality of C-1027 open reading frame polypeptides comprising a

-60-

hydroxylase, a halogenase, an aminomutase, a type II NRPS condensation enzyme, a type II
NRPS adenylation enzyme, and a type II peptidyl carrier protein.

51. A method of synthesizing a chromaprotein type enediyne core, said
5 method comprising contacting a fatty acid with one or more C-1027 orf polypeptides

selected from the group consisting of an epoxide hydrase, a monooxygenase, an iron-sulfer
flavoprotein, a p-450 hydroxylase, an oxidoreductase, and a proline oxidase.

52. The method of claim 51, wherein said fatty acid is contacted with a
plurality of C-1027 orf polypeptides comprising an epoxide hydrase, a monooxygenase, an

10 iron-sulfer flavoprotein, a p-450 hydroxylase, an oxidoreductase, and a proline oxidase.

53. The method of claim 52, wherein said fatty acid is contacted with
polypeptides encoded by ORF17, ORF20, ORF21, ORF29, ORF30, ORF32, ORF35, and
ORF38.

15 contacting a sugar with one or more C-1027 open reading frame polypeptides selected from
the group consisting of a dNDP-glucose synthase, a dNDP glucose dehydratase, an
epimerase, an aminotransferase, a C-methyltransferase, an N-methyltransferase, and a
glycosyl transferase.

55. The method of claim 54, wherein said method comprises contacting a
20 dNDP-glucose with a plurality of C-1027 open reading frame polypeptides comprising a

dNDP-glucose synthase, a dNDP glucose dehydratase, an epimerase, an aminotransferase, a
C-methyltransferase, an N-methyltransferase, and a glycosyl transferase.

56. The method of claim 55, wherein said dNDP-glucose is contacted with
polypeptides encoded by ORF17, ORF20, ORF21, ORF29, ORF30, ORF32, ORF35, and

25 ORF38.

57. A method of synthesizing a beta amino acid, said method comprising
contacting an amino acid with one or one or more C-1027 open reading frame polypeptides
selected from the group consisting of a hydroxylase, an aminomutase, a type II NRPS

50.

The method of claim 48, wherein said amino acid is a tyrosine.

54. A method of synthesizing a deoxysugar, said method comprising

-61-

condensation enzyme, a type II NRPS adenylation enzyme, and a type II peptidyl carrier
protein.

58. The method of claim 57, wherein said method comprises contacting an
amino acid with a plurality of C- 1027 open reading frame polypeptides comprising a

5 hydroxylase, a halogenase, an aminomutase, a type II NRPS condensation enzyme, a type II
NRPS adenylation enzyme, and a type II peptidyl carrier protein.

59. The method of claim wherein said amino acid is contacted with
polypeptides encoded by ORF 4, ORFT 1, ORF24, ORF23, ORF25, and ORF26.

60. The method of claim 57, wherein said amino acid is a tyrosine.

10 6 1 . A method of synthesizing an enediyne or an enediyne analogue said

method comprising:

culturing a cell comprising a recombinantly modified C-1027 gene
cluster under conditions whereby said cell expresses said enediyne or enediyne analogue;
and

15 recovering said enediyne or enediyne analogue.

62. The method of claim 61, wherein said gene cluster is present in a

bacterium.

63. The gene cluster of claim 62, wherein said gene cluster is present in a
bacterium selected from the group consisting of Actinomycetes, Actinoplanetes,

20 Actinomadura, Micrornonospora, and Streptomycetes.

64. The gene cluster of claim 62, wherein said gene cluster is present in a
bacterium selected from the group consisting Streptomyces globisporus, Streptomyces
lividans, Streptomyces coelicolor, Micrornonospora echinospora spp. calichenisis,
Actinomadura verrucosopora, Micrornonospora chersina, Streptomyces carzinostaticus, and

25 Actinomycete L585-6.

65. The method of claim 61, wherein said gene cluster is present in a
eukaryotic cell.

-62-

66. The method of claim 65, wherein said eukaryotic cell is selected from
the group consisting of a mammalian cell, a yeast cell, a plant cell, a fungal cell, and an
insect cell.

67. The method of claim 61, wherein said host cell synthesizes sugars and
5 glycosylates said enediyne or enediyne analogue.

68. The method of claim 67, wherein said host cell synthesizes

deoxy sugars.

69. A method of making a cell resistant to an enediyne or an enediyne
metabolite, said method comprising expressing in said cell one or more isolated C-1027 open

10 reading frame nucleic acids that encode a protein selected from the group consisting of a

CagA apoprotein, a SgcB transmembrane efflux protein, a transmembrane transport protein,
a Na+/H+ transporter, an ABC transport, a glycerol phosphate tranporter, and a UvrA-like
protein.

70. The method of claim 69, wherein said isolated C- 1 027 open reading
15 frame nucleic acids are selected from the group consisting of ORF 9, ORF2, ORF 27, ORF 0,

ORF 1 c-terminus, ORF 2, and ORF 1 N-terminus.

71. The method of claim 69, wherein said cell is a bacterial cell.

-63-

GENE CLUSTER FOR PRODUCTION OF THE ENEDIYNE
ANTITUMOR ANTIBIOTIC C-1027

ABSTRACT OF THE DISCLOSURE

5 This invention provides nucleic acid sequences and characterization of the

gene cluster responsible for the biosynthesis of the enediyne C-1027 (produced by
Streptomyces globisporus). Methods are provided for the biosynthesis of enediynes,
enediyne analogs and other biological molecules.

10
15
20

FILE: c:\_docs\2500 ucott\128usl\2500.128wo0enediyne.apl.doc

-64-

-1-

^OH

^OH

oh|

OP

oh|

ONDP

oh|

ONDP

ORF6

ch^dh]

OH ONDP

ORF8

H 2

OH

ONDP

R-OH

ORF7^ (H 3 C) 2 N^YX1A (H 3 C) 2 N" VU^/OR

CHi)H| [cH^3h] ORF19 |CH£>H

ONDP

OH

ONDP

OH

R = enediyne core

ORF10: dNDP-glucose synthase, 355 aa
ORF1: dNDP-glucose dehydratase, 332 aa
ORF12: epimerase, 192 aa
ORF8: aminotransferase, 410 aa

ORF6: C-methyltransferase, 423 aa
ORF7: N-methyltransferase, 244 aa
ORF19: glycosyl transferase, 459 aa

Fig. 2

-2-

ORF4: Hydroxylase, 527 aa ORF23: Type II NRPS condensation enzyme, 459 aj

ORF11: Hydroxylase/halogenase, 492/494 aa ORF25: Type II NRPS adenylation enzyme, 716 aa
ORF24: Aminomutase, 539 aa ORF26: Type II peptidyl carrier protein, 93 aa

Fig. 3 A

-3-

R - enediyne core

ORF15: Anthranilate synthase I, 493 aa ORF3: Coenzyme F390 synthetase, 463 aa
ORF16: Anthranilate synthase II, 220 aa ORF14: Coenzyme F390 synthetase, 484 aa
ORF28: Q-methyltransferase, 350 aa ORF13: O-acyltransferase, 378 aa

Fig. 3B

-4-

Fatty acid

ORF17: Epoxide hydrase
ORF20: Monooxygenase
ORF21: Iron-sulfur flavoprotein
ORF29: P-450 hydroxylase
ORF30: Oxidoreductase
ORF32: Oxidoreductase
ORF35: Proline oxidase
ORF38: P-450 hydroxylase

ORF13: O-acyltransferase, ORF19:Glycosyl transferase, ORF23: Type II NRPS condensation enzyme

Fig. 4

-5-

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Fig. 7

Gdh

1

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1

SgcA

1

MtmE

1

(-> on S3 pn qi i a
l« Wll o ~ s u o

1

Gdh

58

3 O

SgcA

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59

61

Gdh

118

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no

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118

MtmE

1. -1 urn i i

119

121

Gdh

178

TylA2

178

SgcA

178

MtmE

179

consensus

181

Gdh

238

TylA2

238

SgcA

238

MtmE

239

consensus

241

Gdh

298

EEg0RP^DDg3F: 57

PA^EH^GHgDL: 57

PAgHTsSEjAHSgy : 57

TLT^AE3SDSD0F : 58

NL Va prf: 60

GSWpEd pi PNspY A KAgSDLiAlAyHRThGLdV vTR:180

PEKVgP
PEK^P
PEKVjjP
PEKVijP

PLYGDG
PLYGDG
PLYGDG
PLYGDG

sNNYGp Qf PEKvl PLFiTnllDG VPLYGDG n RdWLHV

AES

GRAG

:237

GAG

grQg

:237

MNS

GRAG

:237

RTG

GRAG

:238

GRaG

:240

NIGGGTEL jN|E
NIGGGTELjjNgE
gjlGGGTEL jNgE

nigggtelSnEe

vLe CG dws

[0QPHT

.lrrB a
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rHryHt

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K

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H

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P

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T

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I

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TKIR^E

3EgV:297
\;R:297
B L:297

3A:298

V DR GHDRR Y S vD TKIr ELGY P :300

jSgJPDGGK :329

RSPgGRELERA : 333
Ie^aHlLDAVG- :332

r3y@AVAA :331

consensus 3 01: f egLA Tv WYrdnRaWWePLk a gg :33 6

TylA2

SgcA

MtmE

298:TGITEl
2 98:VAgED|
299 :ReHgD

-9-

S. globisporus CI 027

B S K

I sgcA I

— * 1 ' -rv

0.25-kb 0.75-kb

X

K/B

pBS1012

(4.3-kb)

2.0-kb

B

B

5. globisporus SB1001 C

K/B

1.0-kb

pBS1012
— \\ —

K

6.3-kb

I

j> — w-

-10-

PATENT APPLICATION DECLARATION

(Attorney's Docket No.: 2500.125US2)

Each of the Applicants named below hereby declares as follows:
L My residence, post office address and country of citizenship given below
are true and correct.

2. I believe I am the original, first and joint inventor of the subject matter
which is claimed and for which a patent is sought in the patent application entitled "GENE
CLUSTERFORPRODUCTION OF THE ENEDIYNE ANTITUMOR ANTIBIOTIC C- 1 027, "

Serial No. , filed January 5, 2000, and I have reviewed and understand the

contents of the specification, including its claims.

3. I acknowledge my duty to disclose to the Office all information known to
me to be material to patentability of this application, in accordance with 37 C.F.R. Section 1.56,
which is defined on the attached page.

I further declare that all statements made herein of my own knowledge are true and
that all statements made on information and belief are believed to be true; and further that these
statements were made with the knowledge that willful false statements and the like so made are
punishable by fine or imprisonment, or both, under Section 1001 of Title 18 of the United States
Code, and that such willful false statements may jeopardize the validity of the application or any
patent issuing thereon.

Date:

Ben Shen
Residence and 1 842 Rushmore Lane
Post Office Address: Davis, California 95616

(Citizenship: People's Republic of China)

Date:

Wen Liu

Residence and Institute of Medicinal Biotechnology
Post Office Address: Tiantan, Beijing, 100005, China

(Citizenship: Peoples Republic of China)

Serial No.: SERIAL NO.

-1-

Date:

Steven D. Christenson
Residence and 1079 Monarch Lane
Post Office Address: Davis, California 95616

(Citizenship: United States)

Date:

Scott Standage
Residence and 63 Tudor Road
Post Office Address: Bornet, Herts, ENS 5NW, United Kingdom
(Citizenship: United Kingdom)

Serial No.: SERIAL NO.

-2-

Section 1.56 Duty to Disclose Information Material to Patentability.

(a) A patent by its very nature is affected with a public interest. The public interest is
best served, and the most effective patent examination occurs when, at the time an application is being
examined, the Office is aware of and evaluates the teachings of all information material to patentability. Each
individual associated with the filing and prosecution of a patent application has a duty of candor and good
faith in dealing with the Office, which includes a duty to disclose to the Office all information known to that
individual to be material to patentability as defined in this section. The duty to disclose information exists
with respect to each pending claim until the claim is cancelled or withdrawn from consideration, or the
application becomes abandoned. Information material to the patentability of a claim that is cancelled or
withdrawn from consideration need not be submitted if the information is not material to the patentability of
any claim remaining under consideration in the application. There is no duty to submit information which
is not material to the patentability of any existing claim. The duty to disclose all information known to be
material to patentability is deemed to be satisfied if all information known to be material to patentability of
any claim issued in a patent was cited by the Office or submitted to the Office in the manner prescribed by
§§ L97(b)-(d) and 1.98. However, no patent will be granted on an application in connection with which
fraud on the Office was practiced or attempted or the duty of disclosure was violated through bad faith or
intentional misconduct. The Office encourages applicants to carefully examine:

(1) prior art cited in search reports of a foreign patent office in a counterpart
application, and

(2) the closest information over which individuals associated with the filing or
prosecution of a patent application believe any pending claim patentably defines, to make sure that
any material information contained therein is disclosed to the Office.

(b) Under this section, information is material to patentability when it is not cumulative
to information already of record or being made of record in the application, and

( 1 ) It establishes, by itself or in combination with other information, a prima facie case
of unpatentability of a claim; or

(2) It refutes, or is inconsistent with, a position the applicant takes in:

(i) Opposing an argument of unpatentability relied on by the Office, or

(ii) Asserting an argument of patentability.

A prima facie case of unpatentability is established when the information compels a conclusion that a claim
is unpatentable under the preponderance of evidence, burden-of-proof standard, giving each term in the claim
its broadest reasonable construction consistent with the specification, and before any consideration is given
to evidence which may be submitted in an attempt to establish a contrary conclusion of patentability.

(c) Individuals associated with the filing or prosecution of a patent application within
the meaning of this section are:

(1) Each inventor named in the application;

(2) Each attorney or agent who prepares or prosecutes the application; and

(3) Every other person who is substantively involved in the preparation or prosecution
of the application and who is associated with the inventor, with the assignee or with anyone to whom
there is an obligation to assign the application.

(d) Individuals other than the attorney, agent or inventor may comply with this section
by disclosing information to the attorney, agent, or inventor.

GENE CLUSTER FOR PRODUCTION OF THE ENEDIYNE
ANTITUMOR ANTIBIOTICC-1027

SEQUENCE LISTING

SEQ ID No. 1. C-1027 gene cluster DNA sequence from 1 to 42,000, ORF-(-7) to ORF-
26

GTCGACTCTAGAGGATCCCGGGTGCGGAGTAGGGGTTACGGACGAAGGAGGGGTGCCCGG

1 + + + + + + 60

CAGCTGAGATCTCCTAGGGCCCACGCCTCATCCCCAATGCCTGCTTCCTCCCCACGGGCC
-7-* *LIGPASYPNRVFSPHG

CGACGCCTGCGGCGAAGGGCGGTTCCTTGAGTTCGAGGCCGGTGGCGAGGACGACGTGGT

61 + + + + + + 120

GCTGCGGACGCCGCTTCCCGCCAAGGAACTCAAGCTCCGGCCACCGCTCCTGCTGCACCA
-7 AVGAAFP PEKLELGTALVVH

CCGCGTCGAGGATCTGCGTGTCGGGGAGCGGCCCAGGGCGCAGCCCCTCGGTCAGGTACG

121 + + + + + + 180

GGCGCAGCTCCTAGACGCACAGCCCCTCGCCGGGTCCCGCGTCGGGGAGCCAGTCCATGC
-7 DADLIQTDPLPGPRLGETLY

GGGTGAGGCCCCTGACGGTCACCTCGAAGCAGCGGTCGTGGGACCGGGCGTCGAGCGCCT

181 + + + + + + 240

CCCACTCCGGGGACTGCCAGTGGAGCTTCGTCGCCAGCACCCTGGCCCGCAGCTCGCGGA
-7 PTLGRVTVEFCRDHSRADLA

CCCCGTCCGCTTCCACAAGGACGACGCCGGGACAGGACTCCCGTGCGGCCTCGACCAGTC

241 + + + + + + 300

GGGGCAGGCGAAGGTGTTCCTGCTGCGGCCCTGTCCTGAGGGCACGCCGGAGCTGGTCAG
-7 EGDAEVLVVGPCSERAAEVL

GGGCGTCGAGGTAGTCCTGGAAGATGCGGCGGGGGGCGGGGCCCTGTTCGGTGAACTTCC

301 + + + + + + 360

CCCGCAGCTCCATCAGGACCTTCTACGCCGCCCCCCGCCCCGGGACAAGCCACTTGAAGG
-7 RADLYDQFIRRPAPGQETFK

ACGAAGCCCAGCGCCGGGGCCAGTCGCGCCGGTCGGCCTCCTGGTTGGCCCAGTTGATGA

361 + + + + + + 420

TGCTTCGGGTCGCGGCCCCGGTCAGCGCGGCCAGCCGGAGGACCAACCGGGTCAACTACT
-7 WSAWRRPWDRRDAEQNAWNI

AGTCGAGCACGTCCTCGCGGAACACCGACATCCTGCCGGCCTGGATATTGAAGACGTGGT

421 + + + + + + 480

TCAGCTCGTGCAGGAGCGCCTTGTGGCTGTAGGACGGCCGGACCTATAACTTCTGCACCA
-7 FDLVDERFVSMRGAQINFVH

CCCAGGGGTTGCCGTCACGGTGATAGGCGACGCCGGCCGAGCGGTAgGCGGCGCGCCGCT

481 + + + + + + 540

GGGTCCCCAACGGCAGTGCCACTATCCGCTGCGGCCGGCTCGCCATcCGCCGCGCGGCGA
-7 DWPNGDRHYAVGASRYAARR

CCAGGAGGACGACTTCCAGCGGTCTTCTCGCGAAATGAAGCAGGCGTATCGCGGTCGCCG

541 + + + + + + 600

GGTCCTCCTGCTGAAGGTCGCCAGAAGAGCGCTTTACTTCGTCCGCATAGCGCCAGCGGC
-7 ELLVVELPRRAFHLLRIATA

TGCCTGCCAGGCCCGCCCCTACGACCAGCACCCTGGGGCGCGCACCCGTCATGCCCATGA

601 + + + + + + 660

ACGGACGGTCCGGGCGGGGATGCTGGTCGTGGGACCCCGCGCGTGGGCAGTACGGGTACT
-7-< TGALGAGVVLVRPRAGTMGM

1

AGCCTCCCCCGCTGACTCAGGGCGgCGCGTCGCGCGCTCCCGTCGGTGTCCTCGCTGACT

661 + + + + + + 720

TCGGAGGGGGCGACTGAGTCCCGCcGCGCAGCGCGCGAGGGCAGCCACAGGAGCGACTGA

GGAAGTTCCCTGACCTGGCGTCAACTCCACTGATCCGTAAGGGGATCGCGGGAGTGGATA

721 + + + + + + 780

CCTTCAAGGGACTGGACCGCAGTTGAGGTGACTAGGCATTCCCCTAGCGCCCTCACCTAT

CGGGTCAGGTCGTGCACGATCGTGGCACCAGACAGATCACCACGTCGATAGGCACTCGTG

781 + + + + + + 840

GCCCAGTCCAGCACGTGCTAGCACCGTGGTCTGTCTAGTGGTGCAGCTATCCGTGAGCAC

AGCCGCGCCCGGGGCTCGACGGGGCGGGGCACCGGCAGGGGCGGCCGCGTGATCAGCCGG

841 + + + + + + 900

TCGGCGCGGGCCCCGAGCTGCCCCGCCCCGTGGCCGTCCCCGCCGGCGCACTAGTCGGCC

AGCCTGTCCGGGGGCGTGCGTGCGGGGCGTCAGCTGTCGATGTCGGGAACGCCAGGGACG

901 + + + + + + 960

TCGGACAGGCCCCCGCACGCACGCCCCGCAGTCGACAGCTACAGCCCTTGCGGTCCCTGC
6-* *SDIDPVGPV-

TCGATCTCGGTGCGGGCGTAGTGGTTGAAGTAGTTGGTGTAGAGGTTCACGGCCACGTGG

961 + + + + + + 1020

AGCTAGAGCCACGCCCGCATCACCAACTTCATCAACCACATCTCCAAGTGCCGGTGCACC
6 D I ETRAYHNFYNTYLNVAVH

ACGAAGACCTCGGCGAGCTCGGTGTCCGTCCATCCCTGTGCCACGGCCGCGTTCCACGAG

1021 4- + + + + + 1080

TGCTTCTGGAGCCGCTCGAGCCACAGGCAGGTAGGGACACGGTGCCGGCGCAAGGTGCTC
6 VFVEALETDTWGQAVAANWS

GCGTCAGACGCCTCGCCCACTTCGCCGGCGATCTCCCTGGCCACCTGGACCAGTGCTTCG

1081 + + + + + + 1140

CGCAGTCTGCGGAGCGGGTGAAGCGGCCGCTAGAGGGACCGGTGGACCTGGTCACGAAGC
6 ADSAEGVEGAIERAVQVLAE-

AGCTTCACGTCGTCGCCGGGCGTCCCCCGGCGAATCGCCACGGTCTCCTCCAGCGTGAAA

1141 + + + + + + 1200

TCGAAGTGCAGCAGCGGCCCGCAGGGGGCCGCTTAGCGGTGCCAGAGGAGGTCGCACTTT
6 LKVDDGPTGRRIAVTEELTF-

CCCGCGACCTTCGCCGACACCGTGTGCGCCGCCTGGCAGTACGCGCACGCGTCGACCGCG

1201 + + + + + + 1260

GGGCGCTGGAAGCGGCTGTGGCACACGCGGCGGACCGTCATGCGCGTGCGCAGCTGGCGC
6 GAVKASVTHAAQCYACADVA

CCCACGGCGAGGGCGATCGCCTCGCGTGTGCGGGCGTCGAACGTTCCATGTTCGGCGACG

1261 + + + + + + 1320

GGGTGCCGCTCCCGCTAGCGGAGCGCACACGCCCGCAGCTTGCAAGGTACAAGCCGCTGC
6 GVALAIAERTRADFTGHEAV

GCTCCGGTGATCGCGGCGTAGGTTTCCAGGACCACGGGGGAATGGGCCATTCCCCCGTGG

1321 + + + + + + 1380

CGAGGCCACTAGCGCCGCATCCAAAGGTCCTGGTGCCCCCTTACCCGGTAAGGGGGCACC
6 AGTIAAYTELVVPSHAMGGH-

ATGTTGAGCACTCGCCCGAACCGCTTCTCCAGTCGGCGCAGGATGTCTCCGCCGGCTGCG

1381 + + + + + + 1440

TACAACTCGTGAGCGGGCTTGGCGAAGAGGTCAGCCGCGTCCTACAGAGGCGGCCGACGC
6 INLVRGFRKELRRLIDGGAA-

GGTGCGGTGTCGATGGTGTGGACGGGAATCCGCGGCATGGGAATGCCTCTCCTCGTAGTG

1441 + + + + + + 1500

CCACGCCACAGCTACCACACCAGCCCTTAGGCGCCGTACCCTTACGGAGAGGAGCATCAC
6-< PATDITHVPIRPM

2

ATGGGAGTTCCTCGTCCCTCCAGTCTGCCCAAGCACCTCCCCCGGTGAGCTGTCCCGGCC

1501 + + + + + + 1560

TACCCTCAAGGAGCAGGGAGGTCAGACGGGTTCGTGGAGGGGGCCACTCGACAGGGCCGG

GCCCTCCGGCCCCTTCTAGGCAGGTCGCCCGGTGGTGCGGCCCCAGGACGTCACCTCGCC

1561 + + + + + + 1620

CGGGAGGCCGGGGAAGATCCGTCCAGCGGGCCACCACGCCGGGGTCCTGCAGTGGAGCGG

GCACCACCGGGAGCCCCGAGGGGCGAGGTCAGAGGCCGAGCACCTCCTCGGCCAGGGCGG

1621 + + + + + + 1680

CGTGGTGGCCCTCGGGGCTCCCCGCTCCAGTCTCCGGCTCGTGGAGGAGCCGGTCCCGCC
-5-* * LGLVEEALA

TGCCCCGAACACGGGCCTCGATCTTGGCGAAGGCCAGGTCGCGTGTGGTGGAGGTGTCGT

1681 + + + + + + 1740

ACGGGGCTTGTGCCCGGAGCTAGAACCGCTTCCGGTCCAGCGCACACCACCTCCACAGCA
-5 TGRVRAE I KAFALDRTTSTD

CGGCGAACGGGGAGAAGCCGCAGTCGTCGCAGGTTCCCAGTTGCTCGACGGGGATGTAGC

1741 + + + + + + 1800

GCCGCTTGCCCCTCTTCGGCGTCAGCAGCGTCCAAGGGTCAACGAGCTGCCCCTACATCG
-5 DAFPSFGCDDCTGLQEVPIY

GGGCGGCGAGCAGGATGCGGTCGCGTACCTGCTCGGGGGTCTCGACCACTGGGTCGATCG

1801 + + + + + + 1860

CCCGCCGCTCGTCCTACGCCAGCGCATGGACGAGCCCCCAGAGCTGGTGACCCAGCTAGC
-5 RAALLIRDRVQEPTEVVPDI

GGTCGGTCACCCCGAGGAAGACGCGGGCGGCAGGGGGCAGGTGGTCACGGACGATGCTCA

1861 + + + + + + 1920

CCAGCCAGTGGGGCTCCTTCTGCGCCCGCCGTCCCCCGTCCACCAGTGCCTGCTACGAGT
-5 PDTVGLFVRAAPPLHDRVIS

GGACCCGCTCGGGGTCCGCTTCGCCGGCCAGTTCGAGATAGAAGTTGCCCGCCTTGAGCT

1921 + + 4- + + + 1980

CCTGGGCGAGCCCCAGGCGAAGCGGCCGGTCAAGCTCTATCTTCAACGGGCGGAACTCGA
-5 LVREPDAEGALELYFNGAKL

GGAAGAGCTTGGGCAGCAGTTCGGCGTAGTCGATGTCGAGGCTGTGCGTGGAGTCCTGGT

1981 + + + + + + 2040

CCTTCTCGAACCCGTCGTCAAGCCGCATCAGCTACAGCTCCGACACGCACCTCAGGACCA
-5 QFLKPLLEAYDIDLSHTSDQ

CGCCGCCGGGGCAGGTGTGTACGCCGATGCGGGCGGTTTCCTCGGCGCTGAAGCGCCCCA

2041 + + + + + + 2100

GCGGCGGCCCCGTCCACACATGCGGCTACGCCCGCCAAAGGAGCCGCGACTTCGCGGGGT
-5 DGGPCTHVGIRATEEASFRG

GGACTTCGTTGTTGAGGGCGATGAAGTCGTCGAGGACGCCGCCGCTGGGGTCGAGCTTGA

2101 + + + + + + 2160

CCTGAAGCAACAACTCCCGCTACTTCAGCAGCTCCTGCGGCGGCGACCCCAGCTCGAACT
-5 LVENNLAI FDDLVGGS PDLK

GGGACAGCCGCCCCTCGGTGAAGTCGAGCTGGACCACGTGTGCCCCCGCGTCCAGGCAGC

2161 + + + + + + 2220

CCCTGTCGGCGGGGAGCCACTTCAGCTCGACCTGGTGCACACGGGGGCGCAGGTCCGTCG
-5 LSLRGETFDLQVVHAGADLC

CTCGGATGTCGGCTTCGGCCTCGTCGGCGAGGTCGCGCAGGAACTGCTCGCGGGGGTAGC

2221 + + + + + + 2280

GAGCCTACAGCCGAAGCCGGAGCAGCCGCTCCAGCGCGTCCTTGACGAGCGCCCCCATCG
-5 GRIDAEAEDALDRLFQERPY

CCTCGATGGGAGTGGCGGGGTAGAGGAGGCTGAGGGCGGAGGGTGCGATGACCGCCTGCT
2281 + + + + + + 2340

3

GGAGCTACCCTCACCGCCCCATCTCCTCCGACTCCCGCCTCCCACGCTACTGGCGGACGA
-5 GE I PTAPYLLSLAS PAIVAQ

TCAGGGGGCGGTCCGTGAGCTGCCGTGCGGCGCGCAGATAGGTTTCGGCCCGCACCTGGT

2341 + + + + + + 2400

AGTCCCCCGCCAGGCACTCGACGGCACGCCGCGCGTCTATCCAAAGCCGGGCGTGGACCA
-5 KLPRDTLQRAARLYTEARVQ

AGCGGAAGGGCCCTTGGGTGATGCTGGGGAGCTGCCGGGTGTGCCCGTCTGCGAAGGGGA

2401 + + + + + + 2460

TCGCCTTCCCGGGAACCCACTACGACCCCTCGACGGCCCACACGGGCAGACGCTTCCCCT
-5 YRFPGQTI SPLQRTHGDAFP

TGACAGCGCCGTCGGGCGAGAGGGTGTCGAGGCCGGTCACGGGGTAGGTGGCGAAGCTCG

2461 + + + + + + 2520

ACTGTCGCGGCAGCCCGCTCTCCCACAGCTCCGGCCAGTGCCCCATCCACCGCTTCGAGC
-5 IVAGDPSLTDLGTVPYTAFS

GCTTGGACTGTTCACCGTCCACGAGGACGGGGCTGCCGACTCGTTCCAGTCGTGTCAGGG

2521 + + + + + + 2580

CGAACCTGACAAGTGGCAGGTGCTCCTGCCCCGACGGCTGAGCAAGGTCAGCACAGTCCC
-5 PKSQEGDVLVPSGVRELRTL

TGTCCGCGACGGCCTGTTCCTGCTGTTTGGCCAGGTCCGTGGCGTCCAGGGTTCCCTGGG

2581 + + + + + + 2640

ACAGGCGCTGCCGGACAAGGACGACAAACCGGTCCAGGCACCGCAGGTCCCAAGGGACCC
-5 TDAVAQEQQKALDTADLTGQ

CATGCGCGGCAAGGGCGTGCAGGAGTGTCGCGGAGCGCGGAAGGCTGCCGATCGGCTCAG

2641 + + + + + + 2700

GTACGCGCCGTTCCCGCACGTCCTCACAGCGCCTCGCGCCTTCCGACGGCTAGCCGAGTC
-5 AHAALAHLLTASRPLSGIPE

TGGCGATGGTCATGGCCGAAGAGTAGGGAAGAGGCTGGGTTTCGAACCACCGCAAAGCTT

2701 + + + + + + 2760

ACCGCTACCAGTACCGGCTTCTCATCCCTTCTCCGACCCAAAGCTTGGTGGCGTTTCGAA
-5-< T A I T M -

TGATTGCCGCTTTTTCAGGGGAAGTTGATGCGAAGTCGCCGAGCGGCGGAACGTGCTGAT

2761 + + + + + + 2820

ACTAACGGCGAAAAAGTCCCCTTCAACTACGCTTCAGCGGCTCGCCGCCTTGCACGACTA

GTATGGGGGGCGGGAGGAGCCTGCGGGGTTCTAGGAGCCGGTCGCGGCCACGGTGGAGGA

2821 + + + + + + 2880

CATACCCCCCGCCCTCCTCGGACGCCCCAAGATCCTCGGCCAGCGCCGGTGCCACCTCCT
-4-* *SGTAAVTSS-

GGTGCCCAGCTGGGAGCGGGGGGTCTTTTCGCCGACGCGGTTGGGCTCGATGGTGCGGGG

2881 + + + + + + 2940

CCACGGGTCGACCCTCGCCCCCCAGAAAAGCGGCTGCGCCAACCCGAGCTACCACGCCCC
-4 TGLQSRPTKEGVRNPEITRP-

GTCGACGGCCTCTCCGGGGGCACCTTGCCGGTAGACGCCTTCGGGGTCGGAGTCCCGGTC

2941 + + + + + + 3000

CAGCTGCCGGAGAGGCCCCCGTGGAACGGCCATCTGCGGAAGCCCCAGCCTCAGGGCCAG
-4 DVAEGPAGQRYVGE PDSDRD-

ATGGGGGAGCAGGAAGAAGACCCGGCGCCGGTACAGACCGCTGTCCGGGTCCGCTTCGGC

3001 + + + + + + 3060

TACCCCCTCGTCCTTCTTCTGGGCCGCGGCCATGTCTGGCGACAGGCCCAGGCGAAGCCG
-4 HPLLFFVRRRYLGSDPDAEA-

GTCGGCCCCGAGTTCGATGTAGCCGATCATGCGGCCGTCGCGGGCGTAGCGCGGCTTGTT

3061 + + + + + + 3120

CAGCCGGGGCTCAAGCTACATCGGCTAGTACGCCGGCAGCGCCCGCATCGCGCCGAACAA
-4 DAGLE IYGIMRGDRAYRPKN-

4

CTTGCGCCGGGGGGTCTTGTCCAGGGCCTGGCGGACGTAGTCGAGTCCCTCGGGATCTTC

3121 + + + + + + 3180

GAACGCGGCCCCCCAGAACAGGTCCCGGACCGCCTGCATCAGCTCAGGGAGCCCTAGAAG
-4 KRRPTKDLAQRVYDLGEPDE-

GAGCCACACGACCTTCGCCTCGTGAACGAGATCGCTGTCGGTCAGTAGCGAGCTCATGGC

3181 + + + + + + 3240

CTCGGTGTGCTGGAAGCGGAGCACTTGCTCTAGCGACAGCCAGTCATCGCTCGAGTACCG
-4-< LWVVKAEHVLDSDTLLS S M -

GGCGACCTCTCCTTCGTCGGCGTGCACCGGGTGGGGAAGCGGTGCCTGCGTGATGTGTGT

3241 + + + + + + 3300

CCGCTGGAGAGGAAGCAGCCGCACGTGGCCCACCCCTTCGCCACGGACGCACTACACACA

TCGTCTGCGGCGGTGGGCCGCAGTGGTGCGGACCGCCCGTGGTGCCGGTTCTCGGCCAAA

3301 + + + + + + 3360

AGCAGACGCCGCCACCCGGCGTCACCACGCCTGGCGGGCACCACGGCCAAGAGCCGGTTT

GCACGGGCAGGTACGTCCTGGGGCACTCACATCGTAGATGGGGTCCGCTTCCGCAGGGCA

3361 + + + + + + 3420

CGTGCCCGTCCATGCAGGACCCCGTGAGTGTAGCATCTACCCCAGGCGAAGGCGTCCCGT

GTGCCTCCGGTCGGAGGACGTTCATTCGTCGGCTGCCAGAGCGAGGTTGGGGTAGAACTT

3421 + + + + + + 3480

CACGGAGGCCAGCCTCCTGCAAGTAAGCAGCCGACGGTCTCGCTCCAACCCCATCTTGAA
-3-* *EDAALALNPYFK-

CCGGCCGTTGGATTTGATCATGTCGGCAGGTGAGGCGAGGCCCACTTCCTGGCGGACCCG

3481 + + + + + + 3540

GGCCGGCAACCTAAACTAGTACAGCCGTCCACTCCGCTCCGGGTGAAGGACCGCCTGGGC
-3 RGNS KIMDAPSALGVEQRVR-

GGTGGCGAAGGCACGGGCGGTCCCGGGGCGGATGCCTTCACTGTGTGCGCACCAGGTGCT

3541 + + + + + + 3600

CCACCGCTTCCGTGCCCGCCAGGGCCCCGCCTACGGAAGTGACACACGCGTGGTCCACGA
-3 TAFARATGPRIGESHACWTS-

GTAGGACGTGTAGAGAAGGCCCTGTTCGACGCGTAGCTCGCTGTTCTCGGGGTCGTGGAG

3601 + + + + + + 3660

CATCCTGCACATCTCTTCCGGGACAAGCTGCGCATCGAGCGACAAGAGCCCCAGCACCTC
-3 YSTYLLGQEVRLESNE PDHL-

GCAGCACTCGGCGAGGAAGCGGCCGATGTGGTCCTCGGTGTTCGCGTATGCGCTGGTGGC

3661 + + + + + + 3720

CGTCGTGAGCCGCTCCTTCGCCGGCTACACCAGGAGCCACAAGCGCATACGCGACCACCG
-3 CCEALFRG I HDE TNAYASTA-

GATGCGGACCCGGTCGGGGCCGGCGAGTGTGTCGCGGGTGGCGAGGTAGCGGCGGGCCCC

3721 + + + + + + 3780

CTACGCCTGGGCCAGCCCCGGCCGCTCACACAGCGCCCACCGCTCCATCGCCGCCCGGGG
-3 I RVRD PGALTDRTALYRRAG-

TTCGGTGAGCCAGTGCAGGATCCCGGGGCCCTCGTCCTGGACGAGTTCGACAGCCAGGTT

3781 + + + + + + 3840

AAGCCACTCGGTCACGTCCTAGGGCCCCGGGAGCAGGACCTGCTCAAGCTGTCGGTCCAA
-3 ETLWHLIGPGEDQVLEVALN-

GTCGATCTTGCGTTCGTCGGGGACGATCCGTTCGAAGGGCAGGAGGCGGATGCGGCGCCA

3841 + + + + + + 3900

CAGCTAGAACGCAAGCAGCCCCTGCTAGGCAAGCTTCCCGTCCTCCGCCTACGCCGCGGT
-3 DIKREDPVIREFPLLRIRRW-

GAAGGCGAAGCCGCCGGTGGAGACCTCGGGGCGGTGGTTGCCCAGCAGCCACAGCTTGTG

3901 + + + + + + 3960

CTTCCGCTTCGGCGGCCACCTCTGGAGCCCCGCCACCAACGGGTCGTCGGTGTCGAACAC

5

-3 FAFGGTSVE PRHNGLLWLKH-

CGTGGGTGTGAAGGAGAAATAGTCCTGCCGCATGCGGCGGGCCTTGATCTTGTCACCGCC

396 1 + + + + + + 4020

GCACCCACACTTCCTCTTTATCAGGACGGCGTACGCCGCCCGGAACTAGAACAGTGGCGG
-3 TPTFS FYDQRMRRAKI KDGG-

GGTCAGCAGGCGGACGCGCGCCTCGTCGAAGCGGTCGTTGGGCTTGAGCTCGCTGCACAC

4021 + + + + + + 4080

CCAGTCGTCCGCCTGCGCGCGGAGCAGCTTCGCCAGCAACCCGAACTCGAGCGACGTGTG
-3 TLLRVRAEDFRDNPKLESCV-

GATGAGGCGGCGGCCGTGGAGTTCGGTGAGCTCGGTGGAGTGTTCGGAGTATGCGCCACG

4081 + + + + + + 4140

CTACTCCGCCGCCGGCACCTCAAGCCACTCGAGCCACCTCACAAGCCTCATACGCGGTGC

-3 I LRRGHLETLETSHESYAGR-

GTCCATGAGGAAACCCGGCGGGGCTGCGTCGGCGTAGTCGCCGAGAATCTGGATCATCAC

4141 + + + + + + 4200

CAGGTACTCCTTTGGGCCGCCCCGACGCAGCCGCATCAGCGGCTCTTAGACCTAGTAGTG
-3 DMLFGPPAADAYDGLIQIMV-

GTCGAGGAGAACGGATTTGCCGTTCTTTCCCTGGCCGTGGAGAAAGGGCAGCACCTGCGC

4201 + + + + + + 4260

CAGCTCCTCTTGCCTAAACGGCAAGAAAGGGACCGGCACCTCTTTCCCGTCGTGGACGCG
-3 DLLVS KGNKGQGHLF P LVQA-

CCCGACGTCACCGGTGATGGAGTAGCCGAGAAGGAGGTGGAGGAAGTCGATCATCTCCCG

4261 + + + + + + 4320

GGGCTGCAGTGGCCACTACCTCATCGGCTCTTCCTCCACCTCCTTCAGCTAGTAGAGGGC
-3 GVDGTI SYGLLLHLFDIMER-

CCCTTCGGCGTCACTGCCGAAGGTGTCTTCGAGGAAACGGTGCCAGCGGGGGGTGGGGAT

4321 + + + + + + 4380

GGGAAGCCGCAGTGACGGCTTCCACAGAAGCTCCTTTGCCACGGTCGCCCCCCACCCCTA
-3 GEADSGFTDELFRHWRPTPI-

GTCCTGGGGGGAGGCGCTGGTGGCGCGGGAGTGGAAGTCCCGGGTGGGGTCGGGCTTGCG

4381 + + + + + + 4440

CAGGACCCCCCTCCGCGACCACCGCGCCCTCACCTTCAGGGCCCACCCCAGCCCGAACGC
-3 DQ P SASTARS HFDRT PD PKR-

CATACGGCCGTTGCGGAGGTCGACCACTCCGTCAGGGGTGCACAGGGCGTAGGGGTCTCC

4441 + + + + + + 4500

GTATGCCGGCAACGCCTCCAGCTGGTGAGGCAGTCCCCACGTGTCCCGCATCCCCAGAGG
-3 MRGNRLDVVGDPTCLAYPDG-

GTCGAGGGTGTCGGGATCGAGGGAGAGGTCGGGAGAGGCCTTTGCCTGGGTGAGGAGCGC

4501 + + + + + + 4560

CAGCTCCCACAGCCCTAGCTCCCTCTCCAGCCCTCTCCGGAAACGGACCCACTCCTCGCG
-3 DLTD PDL S LD P SAKAQTLLA-

CTTCATACCGGTCGTCGACAGGGTGCGGCGTTTGTGGTGGTGCAGTTCCCGGTCGGTGAA

4561 + + + + + + 4620

GAAGTATGGCCAGCAGCTGTCCCACGCCGCAAACACCACCACGTCAAGGGCCAGCCACTT
-3 KMGTTSLTRRKHHHLERDTF-

CAGCCCGCGGGGATCGCTGCCGGGCATCTCCTCCGCCATCTCTCCGGCAGCCCACAGGGC

4621 + + + + + + 4680

GTCGGGCGCCCCTAGCGACGGCCCGTAGAGGAGGCGGTAGAGAGGCCGTCGGGTGTCCCG
-3 LGRPDSGPMEEAMEGAAWLA-

AGCTTTCTCGCCTCCGGCCCGCTTCCACCGGTAGCCGTCCCAGGAGTACCAGCCCAGGCC

4681 + + + + + + 4740

TCGAAAGAGCGGAGGCCGGGCGAAGGTGGCCATCGGCAGGGTCCTCATGGTCGGGTCCGG

6

3 AKEGGARKWRYGDWSYWGLG-

CTCCACGTGCCGGAACTGGTCACGGTAGAGACGGACGAAGAGCTTGGCGTTGCCGCGGTC

4741 + + + + + + 4800

GAGGTGCACGGCCTTGACCAGTGCCATCTCTGCCTGCTTCTCGAACCGCAACGGCGCCAG
3 EVHRFQDRYLRVFLKANGRD-

GGTCAGGCTGGCGGGAATCTCGCCCGCCTCCCAGGCGGTCGCGGCGACGGGGGCCTCGGG

4801 + + + + + + 4860

CCAGTCCGACCGCCCTTAGAGCGGGCGGAGGGTCCGCCAGCGCCGCTGCCCCCGGAGCCC
3 TLSAP I EGAEWATAAVPAEP-

AGCGGCCTGGACAGGGAGGAGCGGCGCTGGGGCCGGGGTGGTTTCGAGGGCCAGCATCTG

4861 + + + + + + 4920

TCGCCGGACCTGTCCCTCCTCGCCGCGACCCCGGCCCCACCAAAGCTCCCGGTCGTAGAC
3 AAQVPLLPAPAPTTELALMQ-

CTGAGCGGCGGCAGTTGCGTCAAAGCGAGGGCCCTCGGCGCTGCTGCTCATGGACGTCCT

4921 + + + + + + 4980

GACTCGCCGCCGTCAACGCAGTTTCGCTCCCGGGAGCCGCGACGACGAGTACCTGCAGGA
3-< QAAATADFRPGEAS S S M -

TCGAGATGGAGCGGTCGGGCGGTCCCCGCTGCGGGAACGGCATGAATGATCTTCCCGGTG

4981 + + + + + + 5040

AGCTCTACCTCGCCAGCCCGCCAGGGGCGACGCCCTTGCCGTACTTACTAGAAGGGCCAC

CGGACAGAGTGCCAGGGGCAGCGCATGTGCGGGGGGACAACGGCCCGTTTCGGACGAGGG

5041 + + + + + + 5100

GCCTGTCTCACGGTCCCCGTCGCGTACACGCCCCCCTGTTGCCGGGCAAAGCCTGCTCCC

CCGGCCGACGGGGGGAAGCAGGGGCCGGCAACCGGGTGGCGGGGCGGCGTGAGCGAGGGC

5101 + + + + + + 5160

GGCCGGCTGCCCCCCTTCGTCCCCGGCCGTTGGCCCACCGCCCCGCCGCACTCGCTCCCG

ACGAGCGGCCCGGTACGGGGGGAAGGGCTCGTCTCTCCGTGGGGCGGCACGTTGTGGTCC

5161 + + + + + + 5220

TGCTCGCCGGGCCATGCCCCCCTTCCCGAGCAGAGAGGCACCCCGCCGTGCAACACCAGG

TCGTCCGTCAGCTTGCGTCTGGCTTCAGCCTCCTGACCCCCAATAAGGCGAAAGCTGCTG

5221 + + + + + + 5280

AGCAGGCAGTCGAACGCAGACCGAAGTCGGAGGACTGGGGGTTATTCCGCTTTCGACGAC

GTCAAGCATCTTTCGTGACACTCGGCGAGGGACTGAAGGGACTGTCTTTCGGAATGAGTG

5281 + + + + + + 5340

CAGTTCGTAGAAAGCACTGTGAGCCGCTCCCTGACTTCCCTGACAGAAAGCCTTACTCAC

TAGGGGGTTGTCGGGTGGGGACCGCGCCTCGACTCCCCGGCGGACGGGATCTGTTCGGTC

5341 + + + + + + 5400

ATCCCCCAACAGCCCACCCCTGGCGCGGAGCTGAGGGGCCGCCTGCCCTAGACAAGCCAG

GGTCCCTTGGGTCCCTCCCCGGATCGCGGCAGGGACCCAAGGGGGCGGTGCGGCGGGCGG

5401 + + + + + + 5460

CCAGGGAACCCAGGGAGGGGCCTAGCGCCGTCCCTGGGTTCCCCCGCCACGCCGCCCGCC

TCGGTGAGGGGCCCCGGTGGAGGGACTGAGGGTCTGTATGGAGCGATAAGAGGGTCTGAA

5461 + + + + + + 5520

AGCCACTCCCCGGGGCCACCTCCCTGACTCCCAGACATACCTCGCTATTCTCCCAGACTT

GGGGCGGAGAGAGTTTCGGTCCCTGCGTTGAGTCCCTGGTCATCACCGCAGGTCAGAGGG

5521 + + + + + + 5580

CCCCGCCTCTCTCAAAGCCAGGGACGCAACTCAGGGACCAGTAGTGGCGTCCAGTCTCCC

GTTTTGAGGGGTGAAAAAGGGACTGAAGGGACTCAACTTCCCCATTATGAGCTGAGTAGA

5581 + + + + + + 5640

CAAAACTCCCCACTTTTTCCCTGACTTCCCTGAGTTGAAGGGGTAATACTCGACTCATCT

7

AGAAAGCAGTATGACGATATCGGCGCCTACATACGCGCGCGTACATAGTGAGCTTATAAT

5641 + + + + + + 5700

TCTTTCGTCATACTGCTATAGCCGCGGATGTATGCGCGCGCATGTATCACTCGAATATTA

GCGGAAGTTGAGTCCCTTCAGTCCCTTTTCGTGGGGTCGTATCCCCTCTGACTGCGTTGA

5701 + + + + + + 5760

CGCCTTCAACTCAGGGAAGTCAGGGAAAAGCACCCCAGCATAGGGGAGACTGACGCAACT

CCGTCGCCGCTCCGCGCAGGGACCGAAGAGGGACCAAGTCCCTGCGCGGGGCGGGCGACG

5761 + + + + + + 5820

GGCAGCGGCGAGGCGCGTCCCTGGCTTCTCCCTGGTTCAGGGACGCGCCCCGCCCGCTGC

GTAATCGTGCAGTGCCCCCTCCCCCGTTTCCCACAGCGAGTCGTCGCTCCCCTGTGAGGC

5821 + + + + + + 5880

CATTAGCACGTCACGGGGGAGGGGGCAAAGGGTGTCGCTCAGCAGCGAGGGGACACTCCG

CGGAGAGGGTCCTAGAACCCCTCAGGGGCCGTTCTGTGGCCCTCTGGGCCTCCTCCTGGC

5881 + + + + + + 5940

GCCTCTCCCAGGATCTTGGGGAGTCCCCGGCAAGACACCGGGAGACCCGGAGGAGGACCG

CATTTACCCCATGGGGGCGCTTGGGGGCGTCAGGAGGGCTTGTGAGGGCTCTGCCGGGAA

5941 + + + + + + 6000

GTAAATGGGGTACCCCCGCGAACCCCCGCAGTCCTCCCGAACACTCCCGAGACGGCCCTT
_ 2 -> MRALPGS-

GTGGCGGATTGCGCATGGCAGGAGATGCCCCGACAGCGGCCGGGAATCGACGATGTCCCC

600 1 + + + + + + 6060

CACCGCCTAACGCGTACCGTCCTCTACGGGGCTGTCGCCGGCCCTTAGCTGCTACAGGGG
-2 GGLRMAGDAPTAAGNRRCPP-

CGACCCCTATCCAGCGTCCGCTGATCCTCAGGAGGCAGACCTTGCAGGCTCCAGAAGCGA

60 61 + + + + + + 6120

GCTGGGGATAGGTCGCAGGCGACTAGGAGTCCTCCGTCTGGAACGTCCGAGGTCTTCGCT
-2 TP I QRPL ILRRQTLQAPEAK-

AGAACGGCCGGTCCCCGGAGCAGCCGCAGGAAGAGCGGATCGTCCTGGACGTATGGCTGG

6121 + + + + + + 6180

TCTTGCCGGCCAGGGGCCTCGTCGGCGTCCTTCTCGCCTAGCAGGACCTGCATACCGACC
-2 NGRSPEQPQEERIVLDVWLA-

CGAACTACCCGTTCCCCACCTATGACGGGCGTGACTTCCTCGCTCCGCTGCGCGAGCGGG

6181 + + + + + + 6240

GCTTGATGGGCAAGGGGTGGATACTGCCCGCACTGAAGGAGCGAGGCGACGCGCTCGCCC
-2 NYPFPTYDGRDFLAPLRERA-

CGGCGGAGTTCGAGCGCGCCCACCCCCGATACCGGGTCGACATCAACGGCCACGACTTCT

6241 + + + + + + 6300

GCCGCCTCAAGCTCGCGCGGGTGGGGGCTATGGCCCAGCTGTAGTTGCCGGTGCTGAAGA
-2 AEFERAHPRYRVDINGHDFW-

GGACCATCCCCGAGAAGGTGGCGCGCGCCACCGCGGAGGGCAGGCCTCCGCACATAGCGG

6301 + + + + + + 6360

CCTGGTAGGGGCTCTTCCACCGCGCGCGGTGGCGCCTCCCGTCCGGAGGCGTGTATCGCC
-2 TIPEKVARATAEGRPPHIAG-

GCTACTACGCCACCGACAGCCAGTTGGCGCGGGACGCGCGCAGGCCCGACGGGAAGCCGG

6361 + + + + + + 6420

CGATGATGCGGTGGCTGTCGGTCAACCGCGCCCTGCGCGCGTCCGGGCTGCCCTTCGGCC
-2 YYATDS QLARDARR PDGKPV-

TCTTCACCTCGGTGGAGGCCGCGTTGGCCGGCCGGACGGAGATACTGGGACACCCGGTGG

6421 + + + + + + 6480

AGAAGTGGAGCCACCTCCGGCGCAACCGGCCGGCCTGCCTCTATGACCCTGTGGGCCACC
-2 FT SVEAALAGRTE I LGH PVV-

TGGTGGAGGACCTCGACCCCGTGGTGCGCGACTCCTACTCGTTCGGGGGCGAGTTGGTGT

8

6481 + + + + + + 6540

ACCACCTCCTGGAGCTGGGGCACCACGCGCTGAGGATGAGCAAGCCCCCGCTCAACCACA
-2 VEDLDPVVRDSYS FGGELVS-

CGCTGCCGCTCACGGTCACCACCATGCTCTGCTACGCCAACTCCTCCCTCCTCGCGCGCG

6541 + + + + + + 6600

GCGACGGCGAGTGCCAGTGGTGGTACGAGACGATGCGGTTGAGGAGGGAGGAGCGCGCGC
-2 LPLTVTTMLCYANS SLLARA-

CCGGTGTTCCGGAGTTGCCCCGTACCTGGGATGAGGTCGAAGCAGCCTGCCAGGCGGTGG

6601 + + + + + + 6660

GGCCACAAGGCCTCAACGGGGCATGGACCCTACTCCAGCTTCGTCGGACGGTCCGCCACC
-2 GVPELPRTWDEVEAACQAVA-

CCAGCGTCGACGGGGGGCCCGGTCACGGAATCACCTGGGCCAACGACGGCTGGGTTTTCC

6661 + + + + + + 6720

GGTCGCAGCTGCCCCCCGGGCCAGTGCCTTAGTGGACCCGGTTGCTGCCGACCCAAAAGG
-2 S VDGG PGHG I TWANDGWV F Q -

AGCAGGCCGTCGCCCTTCAGAACGGGGTGCTGACCGATCAGGACAACGGCCGCTCCGGCT

6721 + + + + + + 6780

TCGTCCGGCAGCGGGAAGTCTTGCCCCACGACTGGCTAGTCCTGTTGCCGGCGAGGCCGA
-2 QAVALQNGVLTDQDNGRS G S -

CCGCCACGACGGTGGACGTCACATCGGACGAGATGCTGGACTGGGTCCGCTGGTGGACGC

6781 + + + + + + 6840

GGCGGTGCTGCCACCTGCAGTGTAGCCTGCTCTACGACCTGACCCAGGCGACCACCTGCG
-2 ATTVDVTSDEMLDWVRWWTH-

ACCTCCATGAGCGCGGCCATTACCTCTACACGGGCGGGCCCTCGGACTGGGGCGGGGCGT

6841 + + + + + + 6900

TGGAGGTACTCGCGCCGGTAATGGAGATGTGCCCGCCCGGGAGCCTGACCCCGCCCCGCA
-2 LHERGHYLYTGG P SDWGGAF-

TCGAGGCTTTCGTCCAGCAGAAGGTCGCATTCACCTTCGACTCGTCCAAGGCCGCCCGGG

6901 + + + + + + 6960

AGCTCCGAAAGCAGGTCGTCTTCCAGCGTAAGTGGAAGCTGAGCAGGTTCCGGCGGGCCC
-2 EAFVQQKVAFTFDS S KAARE-

AACTCATCCAGGCCGGTGCACAGGCCGGTTTCGAGGTCGCGGTGTTCCCGTTGCCCAGGA

6961 + + + + + + 7020

TTGAGTAGGTCCGGCCACGTGTCCGGCCAAAGCTCCAGCGCCACAAGGGCAACGGGTCCT
-2 L I QAGAQAGFEVAVF PL PRN-

ACGCGAAGGCCCCGGTAGCGGGCCAGCCCGTCTCGGGAGACTCCCTGTGGCTGGCCGCGG

7021 + + + + + + 7080

TGCGCTTCCGGGGCCATCGCCCGGTCGGGCAGAGCCCTCTGAGGGACACCGACCGGCGCC
-2 AKAPVAGQPVSGD S LWLAAG-

GACTCGACGAGACCACGCAGGACGGGCTGCTCGCTCTCACCCAGTACCTGATCAGCCCGG

7081 + + + + + + 7140

CTGAGCTGCTCTGGTGCGTCCTGCCCGACGAGCGAGAGTGGGTCATGGACTAGTCGGGCC
-2 LDETTQDGLLALTQYLISPA-

CCAACGCCGCGGACTGGCACCGCACCAACGGTTTCGTACCGGTGACCGGCGCGGCCGGGG

7141 + + + + + + 7200

GGTTGCGGCGCCTGACCGTGGCGTGGTTGCCAAAGCATGGCCACTGGCCGCGCCGGCCCC
-2 NAADWHRTNG FVPVTGAAGE-

AACTGCTGGAAGCGACAGGCTGGTTCGACCGCCGGCCGCAGCAACGGGTGGCCGGGGAGC

7201 + + + + + + 7260

TTGACGACCTTCGCTGTCCGACCAAGCTGGCGGCCGGCGTCGTTGCCCACCGGCCCCTCG
-2 LLEATGWFDRRPQQRVAGEQ-

AGTTGAAGGCGTCCGACCGGTCACCGGCGGCGCTCGGCGCGCTGCTCGGCGACTTCGCGG
7261 + + + + + + 7320

9

TCAACTTCCGCAGGCTGGCCAGTGGCCGCCGCGAGCCGCGCGACGAGCCGCTGAAGCGCC
-2 LKASDRS PAALGALLGDFAA-

CCGTCAACGAGGTCATCACCGCAGCGATGGACGATGTCCTGCGCAGTGGAGCGGACCCCG

7321 + + + + + + 7380

GGCAGTTGCTCCAGTAGTGGCGTCGCTACCTGCTACAGGACGCGTCACCTCGCCTGGGGC
-2 VNEVI TAAMDDVLRSGAD P A -

CGAAGGCCTTCGCCGAAGCCGGCGTGGCCGCCCAGCAACTGCTCGATGCCTACAACGCCC

7381 + + + + + + 7440

GCTTCCGGAAGCGGCTTCGGCCGCACCGGCGGGTCGTTGACGAGCTACGGATGTTGCGGG
-2 KAFAEAGVAAQQLLDAYNAR-

GGAACCGCTCCGGATCCGGGACCCCCTCCGCCGTCTGAGATCCGGTACCGGGGCACAGGG

7441 + + + + + + 7500

CCTTGGCGAGGCCTAGGCCCTGGGGGAGGCGGCAGACTCTAGGCCATGGCCCCGTGTCCC
-2-* NRSGSGTPSAV* -

GCGCCGCCGCCCGCTTTCCCGGCGGGGCACTGGCCGGGGGACATGCTCTCCCGCCCCCGG

7501 + + + + + + 7560

CGCGGCGGCGGGCGAAAGGGCCGCCCCGTGACCGGCCCCCTGTACGAGAGGGCGGGGGCC

CAGGACGTAGGGTCAACCCGCCTGCGCCTTCAGGTGGCGGCGCAGATACTCACCGGTCAG

7561 + + + + + + 7620

GTCCTGCATCCCAGTTGGGCGGACGCGGAAGTCCACCGCCGCGTCTATGAGTGGCCAGTC

* GAQAKLHRRLYEGTL-

GGAGGAATCCGCGGCGAGCAGGTCCTTCGGTGTGCCGGTGAAGACGATCTCGCCGCCCTC

76 21 + + + + + + 7680

CCTCCTTAGGCGCCGCTCGTCCAGGAAGCCACACGGCCACTTCTGCTAGAGCGGCGGGAG
-1 SSDAALLDKPTGTFVI EGGE-

CCGTCCCCCGTCGGGACCCAGGTCGATGATCCAGTCGGCCTGCTGCACCACATCGAGGTT

7681 + + + + + + 7740

GGCAGGGGGCAGCCCTGGGTCCAGCTACTAGGTCAGCCGGACGACGTGGTGTAGCTCCAA
-1 RGGDPGLDI IWDAQQVVDLN-

GTGCTCGATGACCACGACGGTGTTCCCGGCCTCGACGAGCCCGTCCAGGAGCTTCAGCAG

7741 + + + + + + 7800

CACGAGCTACTGGTGCTGCCACAAGGGCCGGAGCTGCTCGGGCAGGTCCTCGAAGTCGTC
-1 HE I VVVTNGAEVLGDLLKLL-

GGTGTCAACGTCCGACATGTGCAGCCCGGTGGTGGGCTCGTCCAGGACATAGACCGTGCC

7801 + + + + + + 7860

CCACAGTTGCAGGCTGTACACGTCGGGCCACCACCCGAGCAGGTCCTGTATCTGGCACGG
-1 TDVDSMHLGTTPEDLVYVTG-

CGTGCGGTGCAGCTGGTCGGCAAGTTTGATCCGCTGCAGTTCACCGCCGGAGAGGCTGGA

7861 + + + + + + 7920

GCACGCCACGTCGACCAGCCGTTCAAACTAGGCGACGTCAAGTGGCGGCCTCTCCGACCT
-1 TRHLQDALKIRQLEGGSLSS-

AAGCGGCTGGCCCAGGCTGAGGTACCCAAGACCGACGTCGACGAGAGCGCGCAGTTTCGG

7921 + + + + + + 7980

TTCGCCGACCGGGTCCGACTCCATGGGTTCTGGCTGCAGCTGCTCTCGCGCGTCAAAGCC
-1 LPQGLS LYGLGVDVLARLKP-

CAGCAGGGCCTTCTCGGTGAAGAACTCGACGGCCTCGTCGGCGGGCAGCTCCAGGACGTC

7981 + + + + + + 8040

GTCGTCCCGGAAGAGCCACTTCTTGAGCTGCCGGAGCAGCCGCCCGTCGAGGTCCTGCAG
-1 LLAKETFFEVAEDAPLELVD-

CGCGATCGACTTCCCGCGAAGCTGGTGCTCCAGGACCTCGGGCTTGAAGCGGCGCCCCTC

8041 + + + + + + 8100

GCGCTAGCTGAAGGGCGCTTCGACCACGAGGTCCTGGAGCCCGAACTTCGCCGCGGGGAG
-1 AISKGRLQHELVEPKFRRGE-

10

ACAGACACCGCAGTGCGTGGTCACCGGATCCATGAAGGCCAGCTCGGTGATGATGACCCC

8101 + + + + + + 8160

TGTCTGTGGCGTCACGCACCAGTGGCCTAGGTACTTCCGGTCGAGCCACTACTACTGGGG
-1 CVGCHTTVPDMFALET I I V G -

GCGGCCCTGGCACTCCTCGCACGACCCCTTGGAGTTGAAGCTGAACAGCGAGGCGTTCGC

8161 + + + + + + 8220

CGCCGGGACCGTGAGGAGCGTGCTGGGGAACCTCAACTTCGACTTGTCGCTCCGCAAGCG
-1 RGQCEECSGKSNFS FLSANA-

GCCGGTCTCCTTCGCGAACAGCTTGCGCAGCGGGTCCATCAGGCCGAGGTAGGAGACCGG

8221 + + + + + + 8280

CGGCCAGAGGAAGCGCTTGTCGAACGCGTCGCCCAGGTAGTCCGGCTCCATCCTCTGGCC
-1 GTEKAFLKRLPDMLGLYSVP-

TGTGGAGCGCGACGAGGCGGCGATCGCGGACTGGTCGACAAAGACCGCGTCGGGGTGCGC

8281 + + + + + + 8340

ACACCTCGCGCTGCTCCGCCGCTAGCGCCTGACCAGCTGTTTCTGGCGCAGCCCCACGCG
-1 TSRS SAAIASQDVFVADPHA-

CTCCATGAATGCCCCGGAGATCAGGCTGCTCTTGCCGGAACCCGCCACCCCGGTCACCGC

8341 + + + + + + 8400

GAGGTACTTACGGGGCCTCTAGTCCGACGAGAACGGCCTTGGGCGGTGGGGCCAGTGGCG
-1 EMFAGS ILSSKGSGAVGTVA-

GGTCAGCACACCGGTGGGCACGGCCACGGAGACCTGCTTCAGGTTGTGGAGATCCGCGTT

8401 + + + + + + 8460

CCAGTCGTGTGGCCACCCGTGCCGGTGCCTCTGGACGAAGTCCAACACCTCTAGGCGCAA
-1 TLVGT PVAVSVQKLNHLDAN-

CTCCACGGTCAGCTCCCCCGTGGGCGGGCGGACCTCCTCCTTCACGCGGGCCCCCCGCCG

8461 + + + + + + 8520

GAGGTGCCAGTCGAGGGGGCACCCGCCCGCCTGGAGGAGGAAGTGCGCCCGGGGGGCGGC
-1 EVTLEGTPPRVEEKVRAGRR-

CAGAGCCTCCCCGGTCCGGGTCTTCGCCTTCCGCAGCTTCGCGAAGGACCCCTCGAACAC

8521 + + + + + + 8580

GTCTCGGAGGGGCCAGGCCCAGAAGCGGAAGGCGTCGAAGCGCTTCCTGGGGAGCTTGTG
-1 LAEGTRTKAKRL KAF S GE FV-

GATCTCGCCCCCGTGCACTCCCGCCCCGGGACCGACATCGACGATGTGGTCGGCGATCTC

8581 + + + + + + 8640

CTAGAGCGGGGGCACGTGAGGGCGGGGCCCTGGCTGTAGCTGCTACACCAGCCGCTAGAG
-1 I EGGHVGAGPGVDVI HDAI E-

GATCACAtcGGGGTCGTGCTCGACGACCAGCACGGTGTTCCCCTTGTCGCGCAGCGCGCG

8641 + + + + + + 8700

CTAGTGTagCCCCAGCACGAGCTGCTGGTCGTGCCACAAGGGGAACAGCGCGTCGCGCGC
-1 I VD PDHEVVLVTNGKDRLAR-

CAGCAGGTCGTTGAGCCGCCCCACGTCGCGCGGGTGCAGGCCGATGCTGGGCTCGTCGAA

8701 + + + + + + 8760

GTCGTCCAGCAACTCGGCGGGGTGCAGCGCGCCCACGTCCGGCTACGACCCGAGCAGCTT
-1 LLDNLRGVDRPHLG I S PEDF-

GATGTACGTGAGCCCGGCCAGACCACTGCCGAGGTGGCGCACCATCTTCAGCCGCTGCCC

8761 + + + + + + 8820

CTACATGCACTCGGGCCGGTCTGGTGACGGCTCCACCGCGTGGTAGAAGTCGGCGACGGG
-1 I YTLGALGS GLHRVMKLRQG-

CTCGCCCCCCGAGAGGTCGGCCGTGGGCCTGTCCAGGGTCAGGTAGCCGAGCCCGATGGA

8821 + + + + + + 8880

GAGCGGGGGGCTCTCCAGCCGGCACCCGGACAGGTCCCAGTCCATCGGCTCGGGCTACCT
-1 EGGSLDATPRDLTLYGLGIS-

11

CACGATCCGCTCCAGGGCCGTGCGCGCGGCTTTCGCGAGAGGGGCAGCGGCCGGCTCCGT

8881 + + + + + + 8940

GTGCTAGGCGAGGTCCCGGCACGCGCGCCGAAAGCGCTCTCCCCGTCGCCGGCCGAGGCA
-1 VI RELATRAAKALPAAAPET-

GACGCCGGCGAGCACCTCCGTGAGGTCGCGGACCTCCATGCTCGAGTAGTCGGCGATGTT

8941 + + + + + + 9000

CTGCGGCCGCTCGTGGAGGCACTCCAGCGCCTGGAGGTACGAGCTCATCAGCCGCTACAA
-1 VGALVETLDRVEMS SYDAIN-

CTTGCCGTCGATCCGGACGTCGAGCGCGGCGGCGTTGAGCCGCGCGCCCCGGCAGGAGGG

9001 + + + + + + 9060

GAACGGCAGCTAGGCCTGCAGCTCGCGCCGCCGCAACTCGGCGCGCGGGGCCGTCCTCCC
-1 KGD I RVDLAAANLRAGRC S P -

ACAGACTCCGTCgGTGACGAAACGTTCGATGACCTCGCGCTTGCGGTCGcTCAGCGCGCT

9061 + + + + + + 9120

TGTCTGAGGCAGcCACTGCTTTGCAAGCTACTGGAGCGCGAACGCCAGCgAGTCGCGCGA
-1 CVGDTVFRE IVERKRDSLAS-

GAGGTCGCGCTTGAGGTTGAgCCGCTCGAACCGGTCGGCcAACCCCTCGTAGTTCGTCTG

9121 + + + + + + 9180

CTCCAGCGCGAACTCCAACTcGGCGAGCTTGGCCAGCCGgTTGGGGAGCATCAAGCAGAC
-1 LDRKLNLRE FRDALGEYNTQ-

GAACTCGGTGCTCTTGGTCTTCAGCGTcACCTTCCCGCCGGTGcCGCGCAGCAGCGTGTC

9181 + + + + + + 9240

CTTGAGCCACGAGAACCAGAAGTCGCAgTGGAAGGGCGGCCACgGCGCGTCGTCGCACAG
-1 FETS KTKLTVKGGTGRLLTD-

CAGCTCCTCGGCGCTGTACTCGGCGATCGGCTTGGCCGGATCCAGACGGCCGGACTTCGC

9241 + + + + + + 9300

GTCGAGGAGCCGCGACATGAGCCGCTAGCCGAACCGGCCTAGGTCTGCCGGCCTGAAGCG
-1 LEEASYEAI PKAPDLRGSKA-

CCAGATCTGCCAGTCCGGGCTACCCACCTTGTACTCGGGGAAAAGGACCGCCCCGTCGTC

9301 + + + + + + 9360

GGTCTAGACGGTCAGGCCCGATGGGTGGAACATGAGCCCCTTTTCCTGGCGGGGCAGCAG
-1 W I QWD PSGVKYE P FLVAGDD-

CAGGGACTTCGAGCGGTCCAGCATCTTGTCCAGGTCGAGGGCGATGCTCTGGCCGAGACC

9361 + + + + + + 9420

GTCCCTGAAGCTCGCCAGGTCGTAGAACAGGTCCAGCTCCCGCTACGAGACCGGCTCTGG
-1 LS KSRDLMKDLDLAI SQGLG-

GTCGCAGTCCGGGCACATGCCCTGGGGGTCGTTGAACGAGAACGCGGAGACGCCGAGCGA

9421 + + + + + + 9480

CAGCGTCAGGCCCGTGTACGGGACCCCCAGCAACTTGCTCTTGCGCCTCTGCGGCTCGCT
-1 DCDPCMGQPDNFS FASVGLS-

GGACGGCCCGTCGTCCTTCGTCGTGCCGAACCGTGCGAACAGGGCCCGGATCATCGGCTG

9481 + + + + + + 9540

CCTGCCGGGCAGCAGGAAGCAGCACGGCTTGGCACGCTTGTCCCGGGCCTAGTAGCCGAC
-1 SPGDDKTTGFRAFLARIMPQ-

TACGTCCGTCATGGTCCCCACCGTGGACCGGGCGTTGCCCCCCACGGGCTTCTGGTCGAC

9541 + + + + + + 9600

ATGCAGGCAGTACCAGGGGTGGCACCTGGCCCGCAACGGGGGGTGCCCGAAGACCAGCTG
-1 VDTMTGVTS RANGGVP KQDV-

GATCACCGGGGTGGTGAGGTTCTCGATCGCCTCGGCCTGAGGACGTTCGTACTTCGGAAG

9601 + + + + + + 9660

CTAGTGGCCCCACCACTCCAAGAGCTAGCGGAGCCGGACTCCTGCAAGCATGAAGCCTTC
-1 IVPTTLNE IAEAQPREYKPL-

CTGGTTGCGGATGTACCAGCTGAAGGTGGAGTTCAGCTGTCGCTGGGCCTCCACGGCCAC

12

9661 + + + + + + 9720

GACCAACGCCTACATGGTCGACTTCCACCTCAAGTCGACAGCGACCCGGAGGTGCCGGTG
-1 QNR I YWS FTSNLQRQAEVAV-

CGTGTCGAAGACGATCGACGACTTGCCCGAACCCGAGACCCCCGTGAAGACCGTGATCTG

9721 + + + + + + 9780

GCACAGCTTCTGCTAGCTGCTGAACGGGCTTGGGCTCTGGGGGCACTTCTGGCACTAGAC
-1 TDFVI SSKGSGSVGTFVTIQ-

GTTGCGGGGAATCGTCAGGGAGACATCTTTGAGGTTGTGGATCCGCGCGCCCGCGATGCG

9781 + + + + + + 9840

CAACGCCCCTTAGCAGTCCCTCTGTAGAAACTCCAACACCTAGGCGCGCGGGCGCTACGC
-1 NRPITLSVDKLNHI RAGAIR-

GATGCCGTCTCCCGGGCCGGATGTTTTTCCCGCGCCGGCGGTGGGGTCGGTGACGCTCAC

984! + + + + + + 9900

CTACGGCAGAGGGCCCGGCCTACAAAAAGGGCGCGGCCGCCACCCCAGCCACTGCGAGTG
-l-< IGDGPGSTKGAGATPDTVSN-

AGAGTTTTCCTCCTGGCTTCCGTACATGATTTACCGTGTCAGCCGGGCAAACCGGCGGAA

9901 + + + + + + 9960

TCTCAAAAGGAGGACCGAAGGCATGTACTAAATGGCACAGTCGGCCCGTTTGGCCGCCTT

CGGTAACCACCTAGCTTGTACTCAGGAGGTGTCCGGGGTCTTCTCCTCCCGTGCTGACTT

9961 + + + + + + 10020

GCCATTGGTGGATCGAACATGAGTCCTCCACAGGCCCCAGAAGAGGAGGGCACGACTGAA
0-* * STDPTKEERASK-

GGGGGCCGGCCCGCCGGACAGGGCCGGCTCCGTGTTCCACCCCGCCAGCCGATCCCCCCG

10021 + + + + + + 10080

CCCCCGGCCGGGCGGCCTGTCCCGGCCGAGGCACAAGGTGGGGCGGTCGGCTAGGGGGGC
0 PAPGGSLAPETNWGALRDGR-

CTCCGTCTCGTCCTCCTCGAGAACGATCCGGCTGCTCGCCCAGCGCAGGATCGGCGGCGC

10081 + + + + + + 10140

GAGGCAGAGCAGGAGGAGCTCTTGCTAGGCCGACGAGCGGGTCGCGTCCTAGCCGCCGCG
0 ETEDEELVIRSSAWRLIPPA-

CGTCACCGAGGTGATGAGGGCGACCAGCACGATGATCGTGAAGGTCACGGTGTCCAGTAC

10141 + + + + + + 10200

GCAGTGGCTCCACTACTCCCGCTGGTCGTGCTACTAGCACTTCCAGTGCCACAGGTCATG
0 TVST I LAVLVI ITFTVTDLV-

GCCGATACGCAGGCCGACCAGGGCGATCACCACCTCGATCATTCCACGCGAGTTCATCCC

10201 + + + + + + 10260

CGGCTATGCGTCCGGCTGGTCCCGCTAGTGGTGGAGCTAGTAAGGTGCGCTCAAGTAGGG
0 G I RLGVLAIVVE IMGRSNMG-

CGCTCCGAGCGCCAGCCCCTCGTAGCGGCTCATCCCGCCACTACGGGCGGCGACGTACGC

10261 + + + + + + 10320

GCGAGGCTCGCGGTCGGGGAGCATCGCCGAGTAGGGCGGTGATGCCCGCCGCTGCATGCG
0 AGLALGEYRSMGGSRAAVYA-

ACCGGCGAACTTGCCGAAAGTGGCCACCAACAGCACCCCGAGGCCCGTGAGCAGCACCGA

10321 + + + + + + 10380

TGGCCGCTTGAACGGCTTTCACCGGTGGTTGTCGTGGGGCTCCGGGCACTCGTCGTGGCT
0 GAFKGFTAVLLVGLGTLLVS-

CGGCTCCGCGAGTGCGGTCAGGTCCATGCGAAGCCCCACACTGCCCAGGAACACCGGTGC

10381 + + + + + + 10440

GCCGAGGCGCTCACGCCAGTCCAGGTACGCTTCGGGGTGTGACGGGTCCTTGTGGCCACG
0 PEALATLDMRLGVSGLFVPA-

GAACACGGCCATGACCAGCGTGCGCAGCGGGGCGAGCCGTACCGGGGCGATGTGCCTCAG

10441 + + + + + + 10500

CTTGTGCCGGTACTGGTCGCACGCGTCGCCCCGCTCGGCATGGCCCCGCTACACGGAGTC

13

0 FVAMVLTRL PALRVPAI HRL-

CAGGGTCGCACCGGCCACGAACGCCCCGAACAACGCCTCCATCCCGGCCGCCGCGGTCT^G

10501 + + + + + + 10560

GTCCCAGCGTGGCCGGTGCTTGCGGGGCTTGTTGCGGAGGTAGGGCCGGCGGCGCCAGTC
0 LTAGAVFAGFLAEMGAAATL-

CGCCCCGTACAGGACGACCACGGCCACGCCGACGGTGACGGCCGATACGGGGACCCGGCT

10561 + + + + + + 10620

GCGGGGCATGTCCTGCTGGTGCCGGTGCGGCTGCCACTGCCGGCTATGCCCCTGGGCCGA
0 AGYLVVVAVGVTVASVPVRS-

GTCACCCGTACGGGACAGCCGCCTGCCGATCGGGCCGCCCACCGCACACGCCGCGGCGAC

10621 + + + + + + 10680

CAGTGGGCATGCCCTGTCGGCGGACGGCTAGCCCGGCGGGTGGCGTGTGCGGCGCCGCTG
0 DGTRSLRRGI PGGVACAAAV-

GAAGACGGTCGTCCAGGCCATCGTGGTCAGGACCACGGGCCCCCCGGCCGCCCCACTCGC

10681 + + + + + + 10740

CTTCTGCCAGCAGGTCCGGTAGCACCAGTCCTGGTGCCCGGGGGGCCGGCGGGGTGAGCG
0 FVTTWAMTTLVVPGGAAGSA-

CAGCGCCGTCACCAGAGCGAGCAGCAGCCAGCCCACCGCGTCGTCGAACACCGCTGCCGC

10741 + + + + + + 10800

GTCGCGGCAGTGGTCTCGCTCGTCGTCGGTCGGGTGGCGCAGCAGCTTGTGGCGACGGCG
0 LATVLALLLWGVADD FVAAA-

GATGAGCAGCTGGCCGACGTTGCGGTGCGTCAGATTCAGGTCGGCGAGCGTCTTGGCGAT

10801 + + + + + + 10860

CTACTCGTCGACCGGCTGCAACGCCACGCAGTCTAAGTCCAGCCGCTCGCAGAACCGCTA
0 ILLQGVNRHTLNLDALTKAI-

CACCGGGAGGGCCGTGACACACATCGCGACCCCGAGGAACAGCGCGAAGACGCCCCGCTC

10861 + + + + + + 10920

GTGGCCCTCCCGGCACTGTGTGTAGCGCTGGGGCTCCTTGTCGCGCTTCTGCGGGGCGAG
0 VPLATVCMAVGL FLAFVGRE-

TCCGGAGTCCGCGAGCAGCGAGGCGGGCACCAGGTAGCCGGTGGCGATGCCCAGCCCCAG

10921 + + + + + + 10980

AGGCCTCAGGCGCTCGTCGCTCCGCCCGTGGTCCATCGGCCACCGCTACGGGTCGGGGTC
0 GSDALLSAPVLYGTAIGLGL-

AGGAATCAGAAGACCCGCCAGGCTGACCCGGGCGGCCAGACCCCCGCGCTTGCGCAGGAT

10981 + + + + + + 11040

TCCTTAGTCTTCTGGGCGGTCCGACTGGGCCCGCCGGTCTGGGGGCGCGAACGCGTCCTA
0 P I LLGALSVRAALGGRKRL I-

CCGGGGGTCGAACTGGGCACCTGCGATGGCCACCAGCAGAAGGACGCCGAACTGGCAGAA

11041 + + + + + + 11100

GGCCCCCAGCTTGACCCGTGGACGCTACCGGTGGTCGTCTTCCTGCGGCTTGACCGTCTT
0 RPDFQAGAIAVLLLVGFQCF-

CGCGTCGAGCAGGTGCGCCTGCGAGATGTCCTCGGGAAACAGCCTGCCGGAAAGTCCCGG

11101 + + + + + + 111^0

GCGCAGCTCGTCCACGCGGACGCTCTACAGGAGCCCTTTGTCGGACGGCCTTTCAGGGCC
0 ADLLHAQS IDEPFLRGSLGP-

CGAGATCTGCCCCAGCAGGGTCGGCCCGAGCAGTACCCCCGCGGTCAGCTCCCCCACCAG

11161 + + + + + + H220

GCTCTAGACGGGGTCGTCCCAGCCGGGCTCGTCATGGGGGCGCCAGTCGAGGGGGTGGTC
0 S I QGLLTPGLLVGATLEGVL-

CGGCGGCAGACCGATCCGGGTCCCCAGCCGTCCCAGACCGTAGGCACAGGCGAGCAGGAG

11221 + + + + + + 11280

GCCGCCGTCTGGCTAGGCCCAGGGGTCGGCAGGGTCTGGCATCCGTGTCCGCTCGTCCTC
0 PPLGIRTGLRGLGYACALLL-

14

GCCGACCTGGAGCAGGAAGACCGTCAGCGGCTCCCCGCCCAGCGGCGACGTGGCTGCGAG

11281 + + + + + + H340

CGGCTGGACCTCGTCCTTCTGGCAGTCGCCGAGGGGCGGGTCGCCGCTGCACCGACGCTC
0 GVQLLFVTLPEGGLPSTAAL-

CACAGCCACGTCAGGACCGCGCACCGGGAACCCAGCCCAGCCCGTCCGTCGACGCGGCCA

11341 + + + + + + 11400

GTGTCGGTGCAGTCCTGGCGCGTGGCCCTTGGGTCGGGTCGGGCAGGCAGCTGCGCCGGT
0-< V A V -

11-* * SRAGPVWGLGDTSAA

GACCCCCCTGCCTCACCGGTCGCTCGGCCCCCGCCTCATCCCCCAGAAGAGCCCGTGCCT

11401 + + + + + + H460

CTGGGGGGACGGAGTGGCCAGCGAGCCGGGGGCGGAGTAGGGGGTCTTCTCGGGCACGGA
11 LGGQRVPREAGAEDGLLARA

GCAGTGCGGCGCTCTGCTCCATGAGGCGGCCCACCACCTTTCCCGGCACGGCGCCGTGCG

11461 + + + + + + 11520

CGTCACGCCGCGAGACGAGGTACTCCGCCGGGTGGTGGAAAGGGCCGTGCCGCGGCACGC
11 QLAASQEMLRGVVKGPVAGH

GCCCGTCGGCGTCGCCCGCAGCGGTGTGCGTCATGCCGGCCATCTCGTCGGACGCCTCGG

11521 + + + + + + H580

CGGGCAGCCGCAGCGGGCGTCGCCACACGCAGTACGGCCGGTAGAGCAGCCTGCGGAGCC
11 PGDADGAATHTMGAMED SAE

AGAACCGCTGCCTGGCCCGGGCCGTGTCGGCGAACTCGTCGGAGGAGACCCCGCCGATCA

11581 + + + + + + 11640

TCTTGGCGACGGACCGGGCCCGGCACAGCCGCTTGAGCAGCCTCCTCTGGGGCGGCTAGT
11 SFRQRARATDAFEDSSVGGI

GTTCGACGAAGGACTGCAGGTCGGAGTCCGCGGTGTTGGAGATCTTCCGGGCCTGCCAGA

11641 + + + + + + II 700

CAAGCTGCTTCCTGACGTCCAGCCTCAGGCGCCACAACCTCTAGAAGGCCCGGACGGTCT

11 LEVFSQLDSDATNS IKRAQW

AATAGGAGTCCTCCGAATGGTGCATGTCGTAGAAGCCGACCAGGAACTCGTAGAAGCGGC

11701 + + + + + + 11760

TTATCCTCAGGAGGCTTACCACGTACAGCATCTTCGGCTGGTCCTTGAGCATCTTCGCCG
11 FYSDESHHMDYFGVLFEYFR

CGTACTCCAGCCGGTAGCGGGCCTCGAACTCCTCGAACGCGCTGGTCTCGTCGACCGACC

11761 + + + + + + 11820

GCATGAGGTCGGCCATCGCCCGGAGCTTGAGGAGCTTGCGCGACCAGAGCAGCTGGCTGG
11 GYELRYRAEFEEFASTEDVS

CGTCCAGGCAGGAGTTGAGCGAGCGCGCTGCCAGCAGTCCGCTGTAGGTGGCGAGGTGCA

11821 + + + + + + H880

GCAGGTCCGTCCTCAACTCGCTCGCGCGACGGTCGTCAGGCGACATCCACCGCTCCACGT
11 GDLCSNLSRAALLGSYTALH

CCCCGGAGGAGAACACCGGGTCGACGAAGCACGCGGCATCCCCGACCAGGGCCATGCCCG

11881 + + + + + + H940

GGGGCCTCCTCTTGTGGCCCAGCTGCTTCGTGCGCCGTAGGGGCTGGTCCCGGTACGGGC
11 VGSSFVPDVFCAADGVLAMG

GCGCCCAGAACTTCGTGTTGCTGTACGACCAGTCCTTGCGGACCCGGAGCTCGCCGTAGG

H941 + + + + + + 12000

CGCGGGTCTTGAAGCACAACGACATGCTGGTCAGGAACGCCTGGGCCTCGAGCGGCATCC
11 PAWFKTNSYSWDKRVRLEGY

GGCCCTCGGTCACCCGGGTGGCCTCGGAGAGCTTCTCCGCGATCAGCGGGCAGGCCGCGA

12001 + + + + + + 12060

CCGGGAGCCAGTGGGCCCACCGGAGCCTCTCGAAGAGGCGCTAGTCGCCCGTCCGGCGCT
11 PGETVRTAESLKEAI LPCAA

15

TGAACGACTCCATCGCCTTCTCGGGGTCGCCCTGCACCAGGCTCGCCGAGTCCCGGTTCA

12061 + + + + + + 12120

11 I FSEMAKEPDGQVLSASDRN

CCACTGCGCCGACACTCGTCAGCTCGGGAGACAGGGGTATGTACCAGAACCACCCGTGCT

12121 + + + + + + 12180

GGTGACGCGGCTGTGAGCAGTCGAGCCCTCTGTCCCCATACATGGTCTTGGTGGGCACGA
11 VVAGVSTLEPSLPIYWFWGH

CGAAGGTGCAGGTGAAGATGTTCCCGGAGTTCGGCTTCGGAAGCCGCTTGCCGCCGTTGA

12181 + + + + + + 12240

GCTTCCACGTCCACTTCTACAAGGGCCTCAAGCCGAAGCCTTCGGCGAACGGCGGCAACT
11 EFTCTFINGSNPKPLRKGGN

AGTAGCCGAACAGGGCCAGGTTGCGGAAGAAGGGCGAGTACTCGCGCTTGGCGCCCGACT

12241 + + + + + + 12300

TCATCGGCTTGTCCCGGTCCAACGCCTTCTTCCCGCTCATGAGCGCGAACCGCGGGCTGA
11 FYGFLALNRFFPSYERKAGS

TCTTGTACAGCCCACCGGTGTTGCCGGAGGCGTCCACGACGAAACGGGAGCCCACCTCGT

12301 + + + + + + 12360

AGAACATGTCGGGTGGCCACAACGGCCTCCGCAGGTGCTGCTTTGCCCTCGGGTGGAGCA
11 KKYLGGTNGSADVVFRS GVE

GCTCGCGCCCCTCGGAGTCCCGGTAGCGCACGCCCCGCACCCGGCCGTCCTCGGCCTTGA

12 361 + + + + + + 12420

CGAGCGCGGGGAGCCTCAGGGCCATCGCGTGCGGGGCGTGGGCCGGCAGGAGCCGGAACT

11 HERGESDRYRVGRVRGDEAK

GCACGTCGAGGACATCGCTGTTCTCCCGCACCTCGACACCGTGCCTGCGAGCGTTGTCGA

12421 + + + + + + 12480

CGTGCAGCTCCTGTAGCGACAAGAGGGCGTGGAGCTGTGGCACGGACGCTCGCAACAGCT
11 LVDLVDSNERVEVGHRRAND

GCAGGATCTGGTCGAACTTCATGCGCTCGACCTGGTACGCGTACCCCGTCGCCCCCGGCA

12481 + + + + + + 12540

CGTCCTAGACCAGCTTGAAGTACGCGAGCTGGACCATGCGCATGGGGCAGCGGGGGCCGT

11 LLIQDFKMREVQYAYGTAGP

TCCGGCGCGAGACGGCGAAGTCGAACGTCCACGGTTCGGGGTTGGCACCCCACTTGAACG

12541 + + + + + + !2600

AGGCCGCGCTCTGCCGCTTCAGCTTGCAGGTGCCAAGCCCCAACCGTGGGGTGAACTTGC
11 MRRSVAFDFTWPEPNAGWKF

TCCCGCCGTGCTTGATCGTGAAGGCTGCCTTCTTCAGCTCGTCGGAGACACCGAGGAGGT

12601 + + + + + + 12660

AGGGCGGCACGAACTAGCACTTCCGACGGAAGAAGTCGAGCAGCCTCTGTGGCTCCTCCA
11 TGGHKITFAAKKLEDSVGLL

GTGCGATGCCGTGGACGGTGGAGGGGAGGAGCGACTCACCGATCTGGTAGCGCGGGAAGG

12 661 + + + + + + 12720

CACGCTACGGCACCTGCCACCTCCCCTCCTCGCTGAGTGGCTAGACCATCGCGCCCTTCC

11 HAIGHVTSPLLSEGIQYRPF

TCTCCTTCTCCAGCTGGAGTACGCGATGGCCCCGCTTGCGGACCAGCGTGGAGACGGTCG

12721 + + + + + + 12780

AGAGGAAGAGGTCGACCTCATGCGCTACCGGGGCGAACGCCTGGTCGCACCTCTGCCAGC
11 TEKELQLVRHGRKRVLTSVT

AGCCCGCCGGACCTCCGCCGACCACGATGACGTCGTACTGCGCTGACACGTCCACGGACT

12781 + + + + + + 12840

TCGGGCGGCCTGGAGGCGGCTGGTGCTACTGCAGCATGACGCGACTGTGCAGGTGCCTGA

ll-< SGAPGGGVVIVDYQASVDM

CTCCTTCTCGCACATCGGGCGTCTCATATTCCCAGGAATCCTCTGGCCCGCCCAGGTGCT

16

12841 + + + + + + 12900

GAGGAAGAGCGTGTAGCCCGCAGAGTATAAGGGTCCTTAGGAGACCGGGCGGGTCCACGA

GCCGCATCTTCGGTATTGCGAAGTCGTGGGCATTCTGCGAGAAGCATGAACCGCGTGGCC

12901 + + + + + + 12960

CGGCGTAGAAGCCATAACGCTTCAGCACCCGTAAGACGCTCTTCGTACTTGGCGCACCGG

CGGTCTACAGTGGCGTGGAATTTCAGTGATTGCGCTGAAGGGCGGCACACGATGAAGGCA

12961 + + + + + + 13020

GCCAGATGTCACCGCACCTTAAAGTCACTAACGCGACTTCCCGCCGTGTGCTACTTCCGT
10-> M K A

CTTGTACTGTCGGGTGGTTCGGGGACCCGCCTGCGCCCGATCAGTTACGCCATGCCGAAG

13021 + + + + + + 13080

GAACATGACAGCCCACCAAGCCCCTGGGCGGACGCGGGCTAGTCAATGCGGTACGGCTTC
10 LVLSGGSGTRLRPI SYAMPK

CAGCTCGTTCCGATCGCCGGGAAGCCAGTCCTTGAATATGTTCTGGATAATATCCGGAAC

13081 + + + + + + 13140

GTCGAGCAAGGCTAGCGGCCCTTCGGTCAGGAACTTATACAAGACCTATTATAGGCCTTG
10 QLVPIAGKPVLEYVLDNIRN

CTCGATATCAAAGAGGTCGCCATTGTCGTCGGTGACTGGGCTCAGGAAATTATTGAGGCA

13141 + + + + + + 13200

GAGCTATAGTTTCTCCAGCGGTAACAGCAGCCACTGACCCGAGTCCTTTAATAACTCCGT
10 LDIKEVAIVVGDWAQEI IEA

ATGGGTGACGGCAGCCGTTTCGGTCTGCGCCTCACCTACATACGCCAGGAGCAACCTCTG

13201 + + + + + + 13260

TACCCACTGCCGTCGGCAAAGC CAGACGCGGAGTGGATGTATGCGGT CCTCGTTGGAGAC
10 MGDGSRFGLRLTYIRQEQPL

GGCATCGCGCACTGCGTGAAACTGGCCCGAGACTTCCTCGACGAGGACGACTTCGTCCTC

13261 + + + + + + 13320

CCGTAGCGCGTGACGCACTTTGACCGGGGTCTGAAGGAGCTGCTCCTGCTGAAGCAGGAG
10 GIAHCVKLARDFLDEDDFVL

TACCTAGGCGACATCATGCTGGACGGAGACCTGTCCGCGCAGGCGGGGCACTTCCTCCAC

13321 + + + + + + 13380

ATGGATCCGCTGTAGTACGACCTGCCTCTGGACAGGCGCGTCCGCCCCGTGAAGGAGGTG
10 YLGDIMLDGDLSAQAGHFLH

ACCCGCCCCGCCGCGCGGATCGTCGTGCGCCAGGTGCCCGACCCCCGGGCCTTCGGGGTG

13381 + + + + - "-- + + 13440

TGGGCGGGGCGGCGCGCCTAGCAGCACGCGGTCCACGGGCTGGGGGCCCGGAAGCCCCAC
10 TRPAARIVVRQVPDPRAFGV

ATCGAGCTGGACGGCGAAGGGCGTGTGCTGCGCCTGGTCGAGAAACCCCGTGAACCGCGC

13441 + + + + + + 13500

TAGCTCGACCTGCCGCTTCCCGCACACGACGCGGACCAGCTCTTTGGGGCACTTGGCGCG
10 IELDGEGRVLRLVEKPREPR

AGCGACCTCGCGGCGGTCGGCGTGTACTTCTTCACCGCGGACGTGCACCGCGCCGTCGAC

13501 + + + + + + 13560

TCGCTGGAGCGCCGCCAGCCGCACATGAAGAAGTGGCGCCTGCACGTGGCGCGGCAGCTG
10 SDLAAVGVYFFTADVHRAVD

GCGATTAGCCCGAGCCGACGGGGCGAGCTGGAAATCACCGACGCCATCCAGTGGCTGCTG

13561 + + + + + + 13620

CGCTAATCGGGCTCGGCTGCCCCGCTCGACCTTTAGTGGCTGCGGTAGGTCACCGACGAC
10 AISPSRRGELEITDAIQWLL

GAGCAGGGCCTGCCGGTCGAGGCCGGCCGCTACACGGACTACTGGAAGGACACCGGCCGG

13621 + + + + + + 13680

CTCGTCCCGGACGGCCAGCTCCGGCCGGCGATGTGCCTGATGACCTTCCTGTGGCCGGCC
10 EQGLPVEAGRYTDYWKDTGR

17

GTCGAGGACGTCGTGGAGTGCAACCGGCGGATGCTCGGCCGTCTGGCGCTCCAGGTGTCG

13681 + + + + + + 13740

CAGCTCCTGCAGCACCTCACGTTGGCCGCCTACGAGCCGGCAGACCGCGAGGTCCACAGC
VEDVVECNRRMLGRLALQVS

GGCGAGGTGGACCCGGAGAGCGAACTGGTGGGTGCGGTGGTCGTCGAGGAGGGCGCCCGG

13741 + + + + + + 13800

CCGCTCCACCTGGGCCTCTCGCTTGACCACCCACGCCACCAGCAGCTCCTCCCGCGGGCC
GEVDPES ELVGAVVVEEGAR

GTGACGCGTTCGCGGGTCGTGGGACCAGCGGTGATCGGCGCGGGCACGGTCGTCGAGGAC

13801 + + + + + + 13860

CACTGCGCAAGCGCCCAGCACCCTGGTCGCCACTAGCCGCGCCCGTGCCAGCAGCTCCTG
VTRSRVVGPAVI GAGTVVED

AGCCAGATCGGACCGTACGCCTCCATCGGCCGGCGCTGCACCGTGCGGGCGTCCCGGCTC

13861 + + + + + + 13920

TCGGTCTAGCCTGGCATGCGGAGGTAGCCGGCCGCGACGTGGCACGCCCGCAGGGCCGAG
SQIGPYAS IGRRCTVRASRL

TCCGACTCCATCGTCCTTGACGACGCCTCGATCCTCGCGGTGAGCGGACTGCACGGCTCG

13921 + + + + + + 13980

AGGCTGAGGTAGCAGGAACTGCTGCGGAGCTAGGAGCGCCACTCGCCTGACGTGCCGAGC
SDS IVLDDAS ILAVSGLHGS

CTGATCGGAAGGGGCGCGCGGATCGCGCCCGGGGCCCGGGGCGAGGCCCGGCACCGGCTG

13981 + + + + + + 14040

GACTAGCCTTCCCCGCGCGCCTAGCGCGGGCCCCGGGCCCCGCTCCGGGCCGTGGCCGAC
L IGRGARIAPGARGEARHRL

GTCGTCGGCGACCACGTGCAGATCGAGATCGCGGCCTGACGCACCCACCGGAGCACCGGG

14041 + + + + + + 14100

CAGCAGCCGCTGGTGCACGTCTAGCTCTAGCGCCGGACTGCGTGGGTGGCCTCGTGGCCC
-* VVGDHVQ I E IAA*-

GGGAGGCTCGGCAGGGGCGTCAGGCCGTAAGAAGGGCTGCCGGGGCGGGACGGACCCGCC

14101 + + + + + + 14160

CCCTCCGAGCCGTCCCCGCAGTCCGGCATTCTTCCCGACGGCCCCGCCCTGCCTGGGCGG

CCGGCAGCCCACAGGTCCCCGGTCCGCGGATATGGGGGACTCGAGGTTCGATCAGCCGAA

14161 + + + + + + 14220

GGCCGTCGGGTGTCCAGGGGCCAGGCGCCTATACCCCCTGAGCTCCAAGCTAGTCGGCTT
* * G F -

GGTCAGAGC CACGTGGCCGAGGTCGAGCC CGGAGTTGCCGGCGCCGAGGTTACAGGCGGC

14221 + + + + + + 14280

CCAGTCTCGGTGCACCGGCTCCAGCTCGGGCCTCAACGGCCGCGGCTCCAATGTCCGCCG
TLAVHGLDLGSNGAGLNCAA-

CGTGGCGCAGTCGACGCTGCCGACCGGCGTGCCTTCGGGCGTGGAGCCCGTGTACGACTT

14281 + + + + + + 14340

GCACCGCGTCAGCTGCGACGGCTGGCCGCACGGAAGCCCGCACCTCGGGCACATGCTGAA
TACDVSGVPTGEPTSGTYSK-

GCGCACGACGAAGCTGAACGACGCCGCTCCGGACGCGTCCGTGGTGAAGGACGTCGCGGT

14341 + + + + + + 14400

CGCGTGCTGCTTCGACTTGCTGCGGCGAGGCCTGCGCAGGCACCACTTCCTGCAGCGCCA
RVVFSFSAAGSADTTFSTAT-

CGCCGGGTTGCACGCGTCCTGGCCACCGACCGGAGCGCACTGGGCGATGTAGTAGGTCTC

14401 + + + + + + 14460

GCGGCCCAACGTGCGCAGGACCGGTGGCTGGCCTCGCGTGACCCGCTACATCATCCAGAG
APNCADQGGVPACQAI YYTE-

GCCGGCGGCGGCACCGCTGACCGACACCGACACGCTCTGTCCGTCACTCAGACCCGAGGC

18

14461 + + + + + + 14520

CGGCCGCCGCCGTGGCGACTGGCTGTGGCTGTGCGAGACAGGCAGTGAGTCTGGGCTCCG
9 GAAAGSVSVSVSQGDSLGSA-

GGGACTGACGGAGAAGGCGGGCGCGGCGAAGGCGACGGACTGTGCGGCGGCGGCCAGGCC

14521 + + + + + + 14580

CCCTGACTGCCTCTTCCGCCCGCGCCGCTTCCGCTGCCTGACACGCCGCCGCCGGTCCGG
9 P S V S FAPAAFAVS QAAAAL G -

GATGGATGCGACGGCCACGACGCCGAACCTGGAAGCACGGCGGGACATGTGACGTAACGA

14581 + + + + + + 14640

CTACCTACGCTGCCGGTGCTGCGGCTTGGACCTTCGTGCCGCCCTGTACACTGCATTGCT
9 I SAVAVVGFRSARRSMHRLS-

CATGCGTAGGCTCCGATTCGAGGAGGGGGTTGATCACTCCATGAAAGGATCACCTCGCCG

14641 + + + + + + I 4700

GTACGCATCCGAGGCTAAGCTCCTCCCCCAACTAGTGAGGTACTTTCCTAGTGGAGCGGC

9-< M -

8-* * R A

GACGGCCGCCTGCATCTCCCTCTGTGCTCTCGTGGATTTCCGGCACGGCACTCCCGTCGA

14701 + + + + + + 14760

CTGCCGGCGGACGTAGAGGGAGACACGAGAGCACCTAAAGGCCGTGCCGTGAGGGCAGCT
8 PRGGADGETSEHIEPVASGD

CGGCCGCCCGCAGAATGCGGCAGACCCCCCGCACCTCCTCCGGCCCCACCGCCGTACCGG

14761 + + + + + + 14820

GCCGGCGGGCGTCTTACGCCGTCTGGGGGGCGTGGAGGAGGCCGGGGTGGCGGCATGGCC
8 VAARLIRCVGRVEEPGVATG

TGGGCAGCGACAGCACCCGCTCGGTGAGCGCCTCCACCTTCGGGAGCGGATCGGGCGCGT

14821 + + + + + + 14880

ACCCGTCGCTGTCGTGGGCGAGCCACTCGCGGAGGTGGAAGCCCTCGCCTAGCCCGCGCA
8 TPLSLVRETLAEVKPLPDPA

GGCGCGCGAGGTCGGACCGGTAGGGCTCGCAGCTGTGGCAGCCGGGGCTGAAGTAGGCGC

14881 + + + + + + 14940

CCGCGCGCTCCAGCCTGGCCATCCCGAGCGTCGACACCGTCGGCCCCGACTTCATCCGCG
8 HRALDSRYPECSHCGPS FYA

GGGCCAGGACGTTGTGCCGTTGGAGCACCGCCTGGAGTTCGTCGCGGTGCAGCCCGGCGC

X4941 + + + + + + 15000

CCCGGTCCTGCAACACGGCAACCTCGTGGCGGACCTCAAGCAGCGCCACGTCGGGCCGCG
8 RALVNHRQLVAQLEDRHLGA

GGACGGCGTCCACCTCGATGACGACGTACTGGCAGTTCGACAGCTCGTTCGGATCCTGCG

15001 + + + + + + 15060

CCTGCCGCAGGTGGAGCTACTGCTGCATGACCGTCAAGCTGTCGAGCAAGCCTAGGACGC
8 RVADVE IVVYQCNSLENPDQ

GGCGGACCCGGACGCCGGGCAGTCCGTCGAGGTACTGCTCGTACAGACGGTAGTTGCGCC

15061 + + + + + + 15120

CCGCCTGGGCCTGCGGCCCGTCAGGCAGCTCCATGACGAGCATGTCTGCCATCAACGCGG
8 PRVRVGPLGDLYQEYLRYNR

GGTTGATCGCGGTGAAGTGATCGGCGGACTCCAGGGAGGTGAGGCCCATGGCCGCGCTGA

15121 + + + + + + 15180

CCAACTAGCGCCACTTCACTAGCCGCCTGAGGTCCCTCCACTCCGGGTACCGGCGCGACT
8 RNIATFHDASELSTLGMAAS

TCTCGTGCATCCGCGCGACCGTTCCGCTCCCGGTGATCTCATGCGCGGCGTTGAGCCCCT

15181 + + + + + + 15240

AGAGCACGTAGGCGCGCTGGCAAGGCGAGGGCCACTAGAGTACGCGCCGCAACTCGGGGA
8 IEHMRAVTGSGTIEHAANLG

GGTGGCGCATGGCCCGGAGCCGGTCGGCCAGGGCGTCGTCGTCGGTGACGATCGCCCCGC

19

15241 + + + + + ^ -j j u w

CCACCGCGTACCGGGCCTCGGCCAGCCGGTCCCGCAGCAGCAGCCACTGCTAGCGGGGCG
QHRMARLRDALADDDTVIAG

CCTCGAAGCTGTTCACGAACTTCGTCGCCTGGAAGCTGAAGATCTCCGCCGTGCCGAAGC

1530 l + + + + + + 15360

GGAGCTTCGACAAGTGCTTGAAGCAGCGGACCTTCGACTTCTAGAGGCGGCACGGCTTCG
GEFSNVFKTAQFSFIEATGF

CGCCGATCGGCTTCGACCGGTAGGTGCAGCCGAAGGCGTGGGCGGCATCGAAGAGCAGGT

15361 + + + + + + 15420

GCGGCTAGCCGAAGCTGGCCATCCACGTCGGCTTCCGCACCCGCCGTAGCTTCTCGTCCA
GGI PKSRYTCGFAHAADFLL

GCAGCCCGTGCTCGGCGGCCAGCTTGGTCAGCTCGTCGATCCGGGCCGGTCTGCCGAAGA

15421 + + + + + + 15480

CGTCGGGCACGAGCCGCCGGTCGAACCAGTCGAGCAGCTAGGCCCGGCCAGACGGCTTCT
HLGHEAALKTLEDIRAPRGF

CGTGCACGTCCAGGATGGCGCGGGTACGCGGGCCGATGAGCCGCTCCACGTGTGCCACGT

15481 + + + + + + 15540

GCACGTGCAGGTCCTACCGCGCCCATGCGCCCGGCTACTCGGCGAGGTGCACACGGTGCA
VHVD L IARTR PG I LREVHAV

CCGCGGTTCCGGTCTCCTCGTCCAGTTCGCAGAAGACAGGCACCGCACCGATCCAGTCCA

15541 + + + + + + 15600

GGCGCCAAGGCCAGAGGAGCAGGTCAAGCGTCTTCTGTCCGTGGCGTGGCTAGGTCAGGT

DATGTEEDLECFVPVAGIWD

GTGCGTGGGCGGTGGCGACCCAGGTGAAGGAGGGCACGATCACCTCGTCCCCAGGACCGA

15601 + + + + + + 15660

CACGCACCCGCCACCGCTGGGTCCACTTCCTCCCGTGCTAGTGGAGCAGGGGTCCTGGCT

LAHATAVWTFS PVIVEDGPG

TGCCCAGGGCCTTCGCGGCGACCTGGATGCCGGTGGTGGCGTTCGATACGGCGACGCAGT

15661 + + + + + + I 5720

ACGGGTCCCGGAAGCGCCGCTGGACCTACGGCCACCACCGCAAGCTATGCCGCTGCGTCA
IGLAKAAVQIGTTANSVAVC

GCCTGACCTGGGTCAGCTCGGCCACACGGGCCTCGAACTCCCGGACCAGGGGGCCGTCAT

15721 + + + + + + 15 780

CGGACTGGACCCAGTCGAGCCGGTGTGCCCGGAGCTTGAGGGCCTGGTCCCCCGGCAGTA
HRVQTLEAVRAEFERVLPGD

TGGTGAACCACAGGCGCTCCAGCGCCCCGTCGATCCGTTCCATCAAACGGTCGCGGGAGC

15781 + + + + + + 15840

ACCACTTGGTGTCCGCGAGGTCGCGGGGCAGCTAGGCAAGGTAGTTTGCCAGCGCCCTCG
NTFWLRELAGDIREMLRDRS

CCACGTTCGGGCGTCCCACGTGCAGCGGTTCGCTGAAGTAGGGCGTGGGTAGGGAGTCCA

15841 + + + + + + 15900

GGTGCAAGCCCGCAGGGTGCACGTCGCCAAGCGACTTCATCCCGCACCCATCCCTCAGGT
GVNPRGVHLPESFYPTPLSD

GACGCACCGGGCCGCCGCTCATGCCGTGCGCACGCCGACGAAGAGGCCGGGGCTGTTGGG

15901 + + + + + + 15 960

CTGCGTGGCCCGGCGGCGAGTACGGCACGCGTGCGGCTGCTTCTCCGGCCCCGACAACCC
DAPGRRSCRAHADEEAGAVG
THRAAAHAVRTPTKRPGLLG -
RTGPPLMPCARRRRGRGCWA-

15901 + + + + + + 15960

* *ATRVGVFLGPSNP-

< LRVPGGSM

CCGGCCGTCGGCCAGCCGGAAGCCGGGCACGAACCGCACCGAGAGCCCCACCGATTCGAA
15961 + + + + + + 16020

20

GGCCGGCAGCCGGTCGGCCTTCGGCCCGTGCTTGGCGTGGCTCTCGGGGTGGCTAAGCTT

7 RGDALRFGPVFRVS LGVSEF-

GGCGTCGGTGTACTGCTCGCGGGTGAAGAGGCTGGAGGTCAGGACCTCGGAGAACTCTCT

16021 + + + + + + 16080

CCGCAGCCACATGACGAGCGCCCACTTCTCCGACCTCCAGTCCTGGAGCCTCTTGAGAGA
7 ADTYQERTFLS STLVESFER-

GAAGCCGGAGGCGTCCGCGACCCGGAACCGGACCTCCAGACGTGACTTGTCGCCCTGGCG

16081 + + + + + + 16140

CTTCGGCCTCCGCAGGCGCTGGGCCTTGGCCTGGAGGTCTGCACTGAACAGCGGGACCGC
7 FGSADAVRFRVELRS KDGQR-

CACGGAGTGCGTCATCCGCGTGATGACACGGCCCTCCTCCTGGTGCAGATGGCCGCCGAC

16141 + + + + + + 16200

GTGCCTCACGCAGTAGGCGCACTACTGTGCCGGGAGGAGGACCACGTCTACCGGCGGCTG
7 VSHTMRTIVRGEEQHLHGGV-

ATGC CCGTCGAGGAAGTTCTCGGGGAAATACCAGGGTTCGGCGACGAGGACTC CCC CGGG

16201 + + + + + + 16260

TACGGGCAGCTCCTTCAAGAGCCCCTTTATGGTCCCAAGCCGCTGCTCCTGAGGGGGCCC
7 HGDLFNEPFYWPEAVLVGGP-

GTTCAGGTGGTGGGCCATGGCCGACACCGCGGCCTTGAGCTCGGTGACGGACCCCATCTC

16261 + + + + + + 16320

CAAGTCCACCACCCGGTACCGGCTGTGGCGCCGGAACTCGAGCCACTGCCTGGGGTAGAG
7 NLHHAMAS VAAKL ETVS GME-

GCCGAGCGCGTTGCCCATGCAGGTGATCGCGTCGAAGGTGCGGCCCAGGTCGAACGAACG

16321 + + + + + + 16380

CGGCTCGCGCAACGGGTACGTCCACTAGCGCAGCTTCCACGCCGGGTCCAGCTTGCTTGC
7 GLANGMCT IADFT RGLDFS R -

CATGTCACCGGCGTGCAGCGGGACGCCGGGAAGCCGGCCCGCCGCCTGCTCCAGCATCGC

16381 + + + + + + 16440

GTACAGTGGCCGCACGTCGCCCTGCGGCCCTTCGGCCGGGCGGCGGACGAGGTCGTAGCG
7 MDGAHL PVGPLRGAAQELMA-

GGGCGCGTACTCGAGGCCCTCCACATGGCCGAAGAGCGTGGCGAGCGTCTCCAGATGGGC

16441 + + + + + + 16500

CCCGCGCATGAGCTCCGGGAGGTGTACCGGCTTCTCGCACCGCTCGCAGAGGTCTACCCG
7 PAYELGEVHGFLTALTELHA-

TCCGGTGCCGCAGGCGACGTCCAGGAGCGACACGGCGTCGGGGCGGGCGGCGAGGATCAG

16501 + + + + + + 16560

AGGCCACGGCGTCCGCTGCAGGTCCTCGCTGTGCCGCAGCCCCGCCCGCCGCTCCTAGTC
7 GTGCAVDLLSVAD PRAAL I L-

CTCGGTGAGCCCGCGGGCCTCCAGGTCGAAGTCCTTGCCGCGGCTGCGGAACACGAGGTC

16561 + + + + + + 16620

GAGCCACTCGGGCGCCCGGAGGTCCAGCTTCAGGAACGGCGCCGACGCCTTGTGCTCCAG
7 ETLGRAELDFDKGRSRFVLD-

GTAGAACTTCGCGTGCTCGGGGCCGTACTCCATCAGACGAGCTCCTTCGCAGACTGGGCG

16621 + + + + + + 16680

CATCTTGAAGCGCACGAGCCCCGGCATGAGGTAGTCTGCTCGAGGAAGCGTCTGACCCGC
7-< YFKAHEPGYEM-

6-* *VLEKASQA-

GAGATGATTCTGGGCTCCGGGATGGGAACGATGAACTTCCCTCCCGCCTCCAGGAAGCGG

16681 + + + + + + 16740

CTCTACTAAGACCCGAGGCCCTACCCTTGCTACTTGAAGGGAGGGCGGAGGTCCTTCGCC
6 SIIRPEPIPVIFKGGAELFR-

CGCTCCTTGCGGACGACCTCGTCGGTGTAGTTCCAGGCGAGGAGGAGGTAGTAGTCCGGC
16741 + + + + + + 16800

21

GCGAGGAACGCCTGCTGGAGCAGCCACATCAAGGTCCGCTCCTCCTCCATCATCAGGCCG
6 REKRVVEDTYNWALLLYYDP-

TCGGTGGCAGCGACCTCCTCCGGAGGAAGGACCGGGATGCGGTTCCCCGGCAGCAGTTTG

16801 + + + + + + 16860

AGCCACCGTCGCTGGAGGAGGCCTCCTTCCTGGCCCTACGCCAAGGGGCCGTCGTCAAAC
6 ETAAVEEPPLVPIRNGPLLK-

CCGTGCTTGAGGCTGGTGGTGTCGCCGCAGACGGTGATGTCCTGATCCGTCAGACCGCAG

16861 + + + + + + 16920

GGCACGAACTCCGACCACCACAGCGGCGTCTGCCACTACAGGACTAGGCAGTCTGGCGTC
6 GHKLSTTDGCVTIDQDTLGC

GCCATCAGCAACTGGGTCCCCTTGGACGGTGCTCCGTAGCCGGCCACGCGGTGGCCGTCC

16921 + + + + + + 16980

CGGTAGTCGTTGACCCAGGGGAACCTGCCACGAGGCATCGGCCGGTGCGCCACCGGCAGG
6 AMLLQTGKS PAGYGAVRHGD

GCGGCCAGACCGCGAACGAGCGTACGGATCGCTTCGGTCACGCGCGTCACCCGCTCGGCG

16981 + + + + + + 17040

CGCCGGTCTGGCGCTTGCTCGCATGCCTAGCGAAGCCAGTGCGCGCAGTGGGCGAGCCGC
6 AALGRVLTRIAETVRTVREA-

AACGCCCGGTAGGGGGCATCCGTCAGCAGTCCGCGCTCCTCCTCCAGGCCGAGCAGCGCC

17041 + + + + + + 17100

TTGCGGGCCATCCCCCGTAGGCAGTCGTCAGGCGCGAGGAGGAGGTCCGGCTCGTCGCGG
6 FARYPADTLLGREEELGLLA-

GCGACCGAGGGCTCCGGGACCCGTGCGGCCGACTCGCGCGCGGCGACGACCGCGATCGAA

17101 + + + + + + 17160

CGCTGGCTCCCGAGGCCCTGGGCACGCCGGCTGAGCGCGCGCCGCTGCTGGCGCTAGCTT
6 AVSPEPVRAASERAAVVAIS-

CCGCCGTGCACGGCGACCCGCTCCACGTCGATGATCCGCAGGCCGTGCGCGCCGAAGAGG

17161 + + + + + + !7220

GGCGGCACGTGCCGCTGGGCGAGGTGCAGCTACTAGGCGTCCGGCACGCGCGGCTTCTCC
6 GGHVAVREVDI IRLGHAGFL-

TGGCGCAGTGTGTGCAGGGAGAAGTACGACAGGTGCTCGTGGTAGATCGTGTCGAACTGG

17221 + + + + + + 17280

ACCGCGTCACACACGTCCCTCTTCATGCTGTCCACGAGCACCATCTAGCACAGCTTGACC
6 HRLTHLSFYSLHEHYITDFQ-

TTCTCGTCGAGCAGGTTCAGCAGGTACGGCACCTCGATGACCAGGACGCCGTCGTCGTCG

17281 + + + + + + 17340

AAGAGCAGCTCGTCCAAGTCGTCCATGCCGTGGAGCTACTGGTCCTGCGGCAGCAGCAGC
6 NEDLLNLLYPVEIVLVGDDD

AGCACTGCGTCGACGCCGTCCAGGATGCGGTGCACGTCGTCGATGTGCGCGAAGCACTGG

17341 + + + + + + 17400

TCGTGACGCAGCTGCGGCAGGTCCTACGCCACGTGCAGCAGCTACACGCGCTTCGTGACC
6 LVADVGDL I RHVDD I HAFCQ-

CGGCCGATGACGGCCTTGGCCCTGCCCTGCTCAAGGGCGATGCGGCCCGCGGGCTCCGGG

17401 + + + + + + 17460

GCCGGCTACTGCCGGAACCGGGACGGGACGAGTTCCCGCTACGCCGGGCGCCCGAGGCCC
6 RGIVAKARGQELAI RGAPE P

CCGAAGAAGTCCGGGTCCGTGGGGATCCCCCGGGCGTTGGCGATCTCGGCGAGGTTGGCC

17461 + + + + + + 17520

GGCTTCTTCAGGCCCAGGCACCCCTAGGGGGCCCGCAACCGCTAGAGCCGCTCCAACCGG
6 GFFDPDTP IGRANAIEALNA-

GCCGGGTCGACCCCGGCCACCCGCATGCCCGCCGCCCGGAACATCGCGAGCTGGGTGCCG

175 2l + + + + + + 17580

CGGCCCAGCTGGGGCCGGTGGGCGTACGGGCGGCGGGCCTTGTAGCGCTCGACCCACGGC

22

6 APDVGAVRMGAARFMALQTG-

ACGTTGCTGCCCAGCTCCACGACCAGGTCGCCGGAGGCGAGGCTTGCCCGGCGGGTCGCC

1758 1 + + + + + + 17640

TGCAACGACGGGTCGAGGTGCTGGTCCAGCGGCCTCCGCTCCGAACGGGCCGCCCAGCGG
6 VNSGLEVVLDGSALSARRTA-

AGCCCGACGATGTGCGCCATGTGCTCGCGGATCTGGTCGGAGTCGGAGGAGACGTAGACG

17641 + + + + + + 17700

TCGGGCTGCTACACGCGGTACACGAGCGCCTAGACCAGCCTCAGCCTCCTCTGCATCTGC
6 LGVIHAMHERIQDSDSSVYV-

TAGTGCTTGAACAGTGTCCCGGGGTCGACGACATGGCGAAGCGTCATCAGCCGGCACGAC

17701 + + + + + + 17760

ATCACGAACTTGTCACAGGGCCCCAGCTGCTGTACCGCTTCGCAGTAGTCGGCCGTGCTG
6 YHKFLTGPDVVHRLTMLRCS-

CGGCACACGATGACGTCGAGCGGGAAGACGTCCTGCGCCTCATCGGCGTCGGCCGGATCG

1776 1 + + + + + + 17820

GCCGTGTGCTACTGCAGCTCGCCCTTCTGCAGGACGCGGAGTAGCCGCAGCCGGCCTAGC
6 RCVIVDLPFVDQAEDADAPD

ACGAACCCGTTGGCCAGCGGCAGCGAGCCGAAGGAGATCACCTCGGTCCAGTCGTCCGCA

17821 -f + + + + + 17880

TGCTTGGGCAACCGGTCGCCGTCGCTCGGCTTCCTCTAGTGGAGCCAGGTCAGCAGGCGT
6 VFGNALPLSGFS IVETWDDA-

CCGCATACACGGCACGTCTCGTCCCGCCTGCATTTCTCCAGCATGAAGTCTCCTGACGGC

17881 + + + + + + 17940

GGCGTATGTGCCGTGCAGAGCAGGGCGGACGTAAAGAGGTCGTACTTCAGAGGACTGCCG
6-< GCVRCTEDRRCKELM-

GAATGCCGACGCATCGGGCCCGTCGGTCCGGGGACGGTCAATCTAGGGTTCCGGCCGACG

17 941 + + + + + + 18000

CTTACGGCTGCGTAGCCCGGGCAGCCAGGCCCCTGCCAGTTAGATCCCAAGGCCGGCTGC

GGCGCTCCACTTCGTATGTGCCCTACTGGTTCAGCGGAGCGGACGGGTGAACGCCCGTAC

18001 + + + + + + 18060

CCGCGAGGTGAAGCATACACGGGATGACCAAGTCGCCTCGCCTGCCCACTTGCGGGCATG
17_* * RL PRT FARV-

GTCCTCGATGAGGAGCTGCGGCTGCTCCATGGCCGCGAAGTGCCCGCCGCGGTCGAACTC

180 61 + + + + + + 18120

CAGGAGCTACTCCTCGACGCCGACGAGGTACCGGCGCTTCACGGGCGGCGCCAGCTTGAG
17 DEILLQPQEMAAFHGGRDFE-

GGTCCACCGCGTCAGGGTCGGCAGGATGCCCTCGGCGAACGACCGGATCGGCCGGGTGGC

18121 + + + + + + 18180

CCAGGTGGCGCAGTCCCAGCCGTCCTACGGGAGCCGCTTGCTGGCCTAGCCGGCCCACCG
17 TWRTLTPLIGEAFSRI PRTA-

GTCGTCCGGGAACACCGCGACGCCGACGGGGGCCGTCAGCGGCCAGGGCCCGCCCCAGGT

1818i + + + + + + 18240

CAGCAGGCCCTTGTGGCGCTGCGGCTGCCCCCGGCAGTCGCCGGTCCCGGGCGGGGTCCA
17 DDPFVAVGVPATLPWPGGWT-

GCGGGCGAAGTCCGCCATGCCGCGAGCCGACTCGTAGTACAACTGAGCGCTGGAACCGGC

18241 + + + + + + 18300

CGCCCGCTTCAGGCGGTACGGCGCTCGGCTGAGCATCATGTTGACTCGCGACCTTGGCCG

17 RAFDAMGRAS EYYLQAS SGA-

CGTCGCGGTCAGCCAGTAGATCATCACGTGGGTGAGCAGCCGGTCCCGGGAGATGGCCTC

18301 + + + + + + 18360

GCAGCGCCAGTCGGTCATCTAGTAGTGCACCCACTCGTCGGCCAGGGCCCTCTACCGGAG
17 TATLWYIMVHTLLRDRS I A E -

23

CTCCACGTTCTTGCCGCCGCTCCACTCCTGGAACTTGTCGAGAATCCAGGCGAGCTGGCC

18361 + + + + + + i0iiZU

GAGGTGCAAGAACGGCGGCGAGGTGAGGACCTTGAACAGCTCTTAGGTCCGCTCGACCGG

17 EVNKGGSWEQFKDL IWALQG-

GACCGGGGAGTCGGTGAGGCCGTAGGCCAGGGTCTGCGGGCGGGTGGCCTGGATGCGCTG

18421 + + + + + + 18480

CTGGCCCCTCAGCCACTCCGGCATCCGGTCCCAGACGCCCGCCCACCGGACCTACGCGAC
17 VPSDTLGYALTQPRTAQ IRQ-

CCAGCCGATGCCGGTGTCGGCGAACTCCCCGCTGTGCGCCAGCTTGCCCAGGTCGCTCTC

18481 + + + + + + 18540

GGTCGGCTACGGCCACAGCCGCTTGAGGGGCGACACGCGGTCGAACGGGTCCAGCGAGAG
17 WGIGTDAFEGSHALKGLDSE-

GTCCAGGCGCCCGATGGCCTCCGGGGCGTCCTGGGGCGGGAAGGTCACCAGCATGTTCAG

18541 + + + + + + 18600

CAGGTCCGCGGGCTACCGGAGGCCCCGCAGGACCCCGCCCTTCCAGTGGTCGTACAAGTC
17 DLRGIAEPADQPPFTVLMNL-

GTGGACGCCGGCCACGTGCTCGGGGTCGGCCAGCCCCAGCTCCAGCGAGACGACCTTTCC

18601 4- 4- + + + + 18660

CACCTGCGGCCGGTGCACGAGCCCCAGCCGGTCGGGGTCGAGGTCGCTCTGCTGGAAAGG
17 HVGAVHE PDALGLELSVVKG-

CCAGTCGCCGCCCTGGGCGACGTAACGCTCGTAGCCGAGGCGGTTCATCAGCTCCGCCCA

1 866 1 + + + + + + 18720

GGTCAGCGGCGGGACCCGCTGCATTGCGAGCATCGGCTCCGCCAAGTAGTCGAGGCGGGT
17 WDGGQAVYREYGLRNMLEAW-

GGCGCGTGCGATCCGCCGCACGTCCCAGCCCGGCTCGGCAGTCGGGCCGGAGAAGCCGTA

18721 + + + + + + 18780

CCGCGCACGCTAGGCGGCGTGCAGGGTCGGGCCGAGCCGTCAGCCCGGCCTCTTCGGCAT
17 ARAI RRVDWGPEATPGS FGY-

GCCCGGCATGGAGGGGACGACGACGTGGAAGGCGTCCGCCGGGTCGCCGCCGTGCGCGCG

1 8781 + + + + + + 18840

CGGGCCGTACCTCCCCTGCTGCTGCACCTTCCGCAGGCGGCCCAGCGGCGGCACGCGCGC
17 GPMS PVVVHFADAPDGGHAR-

CGGGTCGCTCAGCGGCCCGATGACGTCGAGGAACTCGGCGACCGAGCCCGGCCAGCCGTG

18841 + + + + + + 18900

GCCCAGCGAGTCGCCGGGCTACTGCAGCTCCTTGAGCCGCTGGCTCGGGCCGGTCGGCAC
17 PDSLPGIVDLFEAVSGPWGH-

GGTGAGGATCAGCGGGATCGCGTCCGGCTCGGGCGAACGCACGTGAAGGAAGTGCACGTC

18901 + + + + + + 18960

CCACTCCTAGTCGCCCTAGCGCAGGCCGAGCCCGCTTGCGTGCACTTCCTTCACGTGCAG
17 TLILPIADPEPSRVHLFHVD-

GGCGCCGTCGATCGTGGTGACGAACTGGGGGAACGCGTTCAGCTCGGCCTCCGCGGCACG

1896 i + + + + + + 19020

CCGCGGCAGCTAGCACCACTGCTTGACCCCCTTGCGCAAGTCGAGCCGGAGGCGCCGTGC
17 AGD I TTVFQPFANLEAEAAR-

CCAGTCGTAGCCGTGGCGCCAGTGGTCGGTGAGCTCCTTGAGGTAGGACAGCGGCACTCC

19021 + + + + + + 19080

GGTCAGCATCGGCACCGCGGTCACCAGCCACTCGAGGAACTCCATCCTGTCGCCGTGAGG
17 WDYGHRWHDTLEKLY S L PVG-

GCGGTCCCATCCGGATCCGGGTATCTCGGACGGCCACCGGGTCGCGTCGATCCGCCGGGT

19081 + + + + + + 19140

CGQCAGGGTAGGCCTAGGCCCATAGAGCCTGCCGGTGGCCCAGCGCAGCTAGGCGGCCCA
17 RDWGSGPI ESPWRTADIRRT-

TAAGGTCGTCGAATGTCGGACTGGGTCGATCTCGATACGGAAGGGACGCACAGTGAATCC

24

19141 + + + + + + 19200

ATTCCAGCAGCTTACAGCCTGACCCAGCTAGAGCTATGCCTTCCCTGCGTGTCACTTAGG
17-< LTTSHRVPD I E I RFPRM-

ACCCTCGTGATTGTGGGAGCGGGGCGGCGCGAGGCGGCCGCCCCGATGTGATCCGGGGAC

19201 + + + + + + 19260

TGGGAGCACTAACACCCTCGCCCCGCCGCGCTCCGCCGGCGGGGCTACACTAGGCCCCTG

CGTGTCTCAGGCCGGTTCGGCCGGCGCGGCCGCGCCTTCCCGTGCGGAGAAGGACCGCAG

19261 + + + + + + 19320

GCACAGAGTCCGGCCAAGCCGGCCGCGCCGGCGCGGAAGGGCACGCCTCTTCCTGGCGTG
16-* *AP EAPAAAGERAS FSRV-

GGAGGACAGGAAGTTGCGGATCATCGGCATGCCGTGTTCGGTCCGGAAGCTCTCCGGATG

19321 + + + + + + 19380

CCTCCTGTCCTTCAACGCCTAGTAGCCGTACGGCACAAGCCAGGCCTTCGAGAGGCCTAC
16 S SLFNRIMPMGHETRFSEPH-

GAACTGGACGGACTCCACCGGCAGCGAACGGTGGCGCAGGCCCATCACGTACCCGTCGTC

19381 + + + + + + 19440

CTTGACCTGCCTGAGGTGGCCGTCGCTTGCCACCGCGTCCGGGTAGTGCATGGGCAGCAG
16 F QVS EVPLS RHRL GMVYGDD-

CGTGGAGCGCCCGGTGACCTCGAGGGACGGCGGGACCGTGCCCTCCGGCACGATCAGTGA

19441 + + + + + + 19500

GCACCTCGCGGGCCACTGGAGCTCCCTGCCGCCCTGGCACGGGAGGCCGTGCTAGTCACT
16 TSRGTVELSPPVTGEPVILS-

GTGGTAGCGGGTCGCGAAGAACCCCGCGGGCAGCCCGGTGAACACTCCGCGCCCGTCGTG

19501 + + + + + + 19560

CACCATCGCCCAGCGCTTCTTGGGGCGCCCGTCGGGCCACTTGTGAGGCGCGGGCAGCAC
16 HYRTAFFGAPLGTFVGRGDH-

CGTGATCCGGCTCGTCTTCCCGTGCATGAGATGCCGGGCGGGGACGGTGGCGGCGCCGTA

19561 + + + + + + 19620

GCACTAGGCCGAGCAGAAGGGCACGTACTCTACGGCCCGCCCCTGCCACCGCCGCGGCAT
16 TIRSTKGHMLHRAPVTAAGY-

GGCGCGGGCGACGGCCTGATGCCCCAGACAGACCCCGAGCAGCGGGACCCGGCCGGCGAA

19621 + + + + + + 19680

CCGCGCCCGCTGCCGGACTACGGGGTCTGTCTGGGGCTCGTCGCCCTGGGCCGGCCGCTT
16 ARAVAQHGIiCVGLL PVRGAF-

GGCCTGGACGATCTCGACGTGCCCGGAGGTGTCGGGGTGGCCGGGGCCCGGCCCCAGCAG

19681 + + + + + + 19740

CCGGACCTGCTAGAGCTGCACGGGCCTCCACAGCCCCACCGGCCCCGGGCCGGGGTCGTC
16 AQVI EVHGSTDPHGPGPGLL-

GACCGCGTCCGGCCGCATCAGCCCCATCTCGTCCGGGGTCATGAGATGCGACCGCACCAT

19741 + + + + + + 19800

CTGGCGCAGGCCGGCGTAGTCGGGGTAGAGCAGGCCCCAGTACTCTACGCTGGCGTGGTA
16 VAD PRMLGMED P TMLH S RVM-

GACGGGCTCCGCGCCGGCGGACATCAGATACTGGCGCAGGATGTCGACGAAGCTGTCGAA

19801 + + + + + + 19860

CTGCCCGAGGCGCGGCCGCCTGTAGTCTATGACCGCGTCCTACAGCTGCTTCGACAGCTT
16 VPEAGASMLYQRLIDVFSDF-

CGCGTCGACCACCAGGACCCGCGGGGCCTCGGTGCCTGCGCCGGATCCGTCGGGAGACCA

19861 + + + + + + 19920

GCGCAGCTGGTGGTCCTGGGCGCCCCGGAGCCACGGACGCGGCCTAGGCAGCCCTCTGGT
16 ADVVLVRPAETGAGSGDPSW-

CAAGCTCACAGCAACTCCTCTCCGGTGACCGCCCAGTGAGTGGCGCTCATCTTGGCCAGC

19921 + + + + + + 19980

GTTCGAGTGTCGTTGAGGAGAGGCCACTGGCGGGTCACTCACCGCGAGTAGAACCGGTCG

25

16-< L S M -

15-* * LLEEGTVAWHTASMKAL

GTCTCGGTCCACTCCGCCCCCGGTTCGGAATCGGCGACGATTCCGGCCGAGGCCCGGGTG

19981 + + + + + + 20040

CAGAGCCAGGTGAGGCGGGGGCCAAGCCTTAGCCGCTGCTAAGGCCGGCTCCGGGCCCAC
15 TETWEAGPESDAVIGASART-

CGGTAGACGCCCTCGTGGTGGAAAAGGGTCCGGATGCACAGCGCGAGGTTGGTGTACCCG

20041 + + + + + + 20100

GCCATCTGCGGGAGCACCACCTTTTCCCAGGCCTACGTGTCGCGCTCCAACCACATGGGC
15 RYVGEHHFLTRI CLALNTYG

CCCACGTCGAGGAGGCCGAGCGCCCCGGCGTACAGGCCGCGGCGGCTGCGTTCGACGGAC

20101 + + + + + + 20160

GGGTGCAGCTCCTCCGGCTCGCGGGGCCGCATGTCCGGCGCCGCCGACGCAAGCTGCCTG
15 GVDLLGLAGAYLGRRSREVS-

TCGATGATCTCCATGGCGCGGATCTTCGGCGCGCCCGTCATGGTGCCGGCGGGGAACAGG

20161 + + + + + + 20220

AGCTACTAGAGGTACCGCGCCTAGAAGCCGCGCGGGCAGTACCACGGCCGCCCCTTGTCC
15 EI IEMARI KPAGTMTGAPFL

GCGGCGATGGTGTCGAAGGCATCGGTGTCCACCCGCGCCCGGCCGACGACCGTGGAGACC

20221 + + + + + + 20280

CGCCGCTACCACAGCTTCCGTAGCCACAGGTGGGCGCGGGCCGGCTGCTGGCACCTCTGG
15 AAITDFADTDVRARGVVTSV-

AGGTGCAGCACGTGGGAGTAGCCCTCCACGTCCAGCTGGTCGGGTACGTCGAGCGTGTTC

20281 + + + + + + 20340

TCCACGTCGTGCACCCTCATCGGGAGGTGCAGGTCGACCAGCCCATGCAGCTCGCACAAG
15 LHLVHSYGEVDLQDPVDLTN

GGCCGGGCGATCCGTCCGATGTCGTTGCGGCAGAGGTCCACCAGCATGGTGTGCTCGGCG

20341 + + + + + + 20400

CCGGCCCGCTAGGCAGGCTACAGCAACGCCGTCTCCAGGTGGTCGTACCACACGAGCCGC
15 PRAIRGIDNRCLDVLMTHEA-

ATCTCCTTGGGATCCGACCTCAGCCGGACTCCCGCGGCGATGCCGCCGTCCGCGCCGGAC

20401 + + + + + + 20460

TAGAGGAAGCCTAGGCTGGAGTCGGCCTGAGGGCGCCGCTACGGCGGCAGGCGCGGCCTG
15 IEKPDSRLRVGAAIGGDAGS

CGCGGCACCGTGCCCGCGATCGGCCGCATCGTGACCTCGCCGTCCTCGATGCGTACGAAC

20461 + + + + + + 20520

GCGCCGTGGCACGGGCGCTAGCCGGCGTAGCACTGGAGCGGCAGGAGCTACGCATGCTTG
15 RPVTGAI PRMTVEGDE I RVF

AGCTCGGGGCTGGCGCCGATCAGACGGTGCCCGTCGATGCCCGCCAGATACATGTACGGG

20521 + + + + + + 20580

TCGAGCCCCGAGCGCGGCTAGTCTGCCACGGGCAGCTACGGGCGGTCTATGTACATGCCC
15 LEPSAGILRHGDIGALYMYP-

GAGGCGTTCCGCCCGCGCAGGCGCTGGTAGACGTCCGCGGGGTCGGCCGTCGAGCGGATG

20581 + + + + + + 20640

CTCCGCAAGGCGGGCGCGTCCGCGACCATCTGCAGGCGCCCCAGCCGGCAGCTCGCCTAC
15 SANRGRLRQYVDAPDATSRI-

GAGAGCTCGTGAC CGATCTGCACCTGGTAGATGTCGC CGACGGCGATGTGCTTCAGACAC

20641 + + + + + + 20700

CTCTCGAGCACTGGCTAGACGTGGACCATCTACAGCGGCTGCCGCTACACGAAGTCTGTG
15 SLEHGIQVQYIDGVAIHKLC

CGCTCGACGTCGTTCGCGAACACTTCGGGGGCGCTGTCGTCGGTGACCGCGGAGGCGGGG

20701 + + + + + + 20760

GCGAGCTGCAGCAAGCGCTTGTGAAGCCCCCGCGACAGCAGCCACTGGCGCCTCCGCCCC

26

15 REVDNAFVEPASDDTVASAP

AAGCCGTCTGCGGACGGATCGGGCCAGGCCTGCTCCACGTCGGCGAGGAGCCCGGTGACG

20761 + + + + + + 20820

TTCGGCAGACGCCTGCCTAGCCCGGTCCGGACGAGGTGCAGCCGCTCCTCGGGCCACTGC

15 FGDAS PD PWAQEVDALLGTV

GTCTCCGGCGCGAGGCCGGGCCAGTACGGGGACTCGTGGAGCAGCAGTTCGCATCGGCCG

20821 + + + + + + 20880

CAGAGGCCGCGCTCCGGCCCGGTCATGCCCCTGAGCACCTCGTCGTCAAGCGTAGCCGGC
15 TEPALGPWYPSEHLLLECRG-

GTGGCGAGATCGGTGACCACGCTGCCCCGGTGCAGGACCATGCGTACGTCCGGCAGGCCA

20881 + + + + + + 20940

CACCGCTCTAGCCACTGGTGCGACGGGGCCACGTCCTGGTACGCATGCAGGCCGTCCGGT

15 TALDTVVSGRHLVMRVDPLG-

GGCCGGTTCTCGATGAGGTGGGGCAGGTCCTCGATGTAGCGGGCCGTGTCGTACCCGAAG

20941 + + + + + + 21000

CCGGCCAAGAGCTACTCCACCCCGTCCAGGAGCTACATCGCCCGGCACAGCATGGGCTTC
15 PRNEILHPLDEIYRATDYGF

AACCCGAGGAACCCGAAGCGGAAGCCGGACGCGGACCCCTCGGCGTCGAACATGTCCCGC

21001 + + + + + + 21060

TTGGGCTCCTTGGGCTTCGCCTTCGGCCTGCGCCTGGGGAGCCGCAGCTTGTACAGGGCG
15 FGLFGFRFGSASGEADFMDR

ATGGCCCGCAGCAGCGGCCACAACCCGCCCGCGGTACGCAGCCGCAGCCCCTGGGGGCCG

21061 + + + + + + 21120

TACCGGGCGTCGTCGCCGGTGTTGGGCGGGCGCCATGCGTCGGCGTCGGGGACCCCCGGC
15 MARLLPWLGGATRLRLGQPG-

TCCTCCAGGAGCGCGCCGGCCCGCTCCAGGAGCAGGCCCCGCAGGGCGGGTACGCCCTCG

21121 + + + + + + 21180

AGGAGGTCCTCGCGCGGCCGGGCGAGGTCCTCGTCCGGGGCGTCCCGCCCATGCGGGAGC
15 DELLAGARELLLGRLAPVGE-

ACGCGCACCACCCGGTCGGTGACCGAGAGCGAGAGCAGCGCGCCGAAGCCGACGAACTGG

2H81 + + + + + + 21240

TGCGCGTGGTGGGCCAGCCACTGGCTCTCGCTCTCGTCGCGCGGCTTCGGCTGCTTGACC
15 VRVVRDTVSLSLLAGFGVFQ

TGCCTGCGGTCGCGGGCCGGGCCGGCCGCGGACTCCAGGAGGTAGACCTCGTCGGGGCCG

21241 + + + - + + + 21300

ACGGACGCCAGCGCCCGGCCCGGCCGGCGCCTGAGGTCCTCCATCTGGAGCAGCCCCGGC
15 HRRDRAPGAASELLYVEDPG-

AAGTGCTCGGCCAGCGCGCGGTAGGCGGGCAGGGCGCCCGTCTCCTTCACATCGAGGCGT

21301 + + + + + + 21360

TTCACGAGCCGGTCGCGCGCCATCCGCCCGTCCCGCGGGCAGAGGAAGTGTAGCTCCGCA
15 FHEALARYAPLAGTEKVDLR

CGTGTCCGCACCCGCACCGGGGCCGAGACCACGCACTGGTCGGTCATCCTGGGTCCTCCC

21361 + + + + + + 21420

GCACAGGCGTGGGCGTGGCCCCGGCTCTGGTGCGTGACCAGCCAGTAGGACCCAGGAGGG
15-< RTRVRVPASVVCQDTM-

GGATCACGTGGTGATGGCGTAGCGGTGTGCCACCTGACGGGCGGTCAGCACCGCCCGGTC

21421 + + + + + + 21480

CCTAGTGCACCACTACCGCATCGCCACACGGTGGACTGCCCGCCAGTCGTGGCGGGCCAG

14-* * TTIAYRHAVQRATLVARD-

GGGGCCGGAGCGGTTGTCGACGACGCGCGCGGCCTTCCAGCTGACGAAGGAGCCGGTGTG

21481 + + + + + + 21540

CCCCGGCCTCGCCAACAGCTGCTGCGCGCGCCGGAAGGTCGACTGCTTCCTCGGCCACAC
14 PGSRNDVVRAAKWSVFSGTH-

27

GGTCACGGGGTCGAGGTCGGTGTCCACGACGATGCCGGCGTGCGCGCCGGTCCGCTCCCT

21541 + + + + + + 21600

CCAGTGCCCCAGCTCCAGCCACAGGTGCTGCTACGGCCGCACGCGCGGCCAGGCGAGGGA
14 TVPDLDTDVV I GAHAGTRER-

GAGCCGGGCGGCGACGGCCTCGCCGATGCCCTGCCGTTCCCCCTCGGCGCCGGCCAGCAG

21601 + + + + + + 21660

CTCGGCCCGCCGCTGCCGGAGCGGCTACGGGACGGCAAGGGGGAGCCGCGGCCGGTCGTC
14 LRAAVAEG I GQREGEAGALL-

GTCCATGCGCACGGTGACGGCGTCGCTGCCGTCGTCCTGCCGGTCGATGACGACCTGGTA

21661 + + + + + + 21720

CAGGTACGCGTGCCACTGCCGCAGCGACGGCAGCAGGACGGCCAGCTACTGCTGGACCAT
14 DMRVTVADSGDDQRDIVVQY-

GCCGAGGCAGCCGCCGACCCCGTCGAGGATCGCGGCCTCCAGCTCGGCGGGCTGGAGGGT

21721 + + + + + + 21780

CGGCTCCGTCGGCGGCTGGGGCAGCTCCTAGCGCCGGAGGTCGAGCCGCCCGACCTCCCA
14 GLCGGVGDLIAAELEAPQLT-

CACGTCGCCCAGGGGGATGCGGTCCGCGACCCGGCCGATGACCTGGATCCGCGGTCCCGG

21781 + + + + + + 21840

GTGCAGCGGGTCCCCCTACGCCAGGCGCTGGGCCGGCTACTGGACCTAGGCGCCAGGGCC
14 VDGLP I RDAVRG I VQ I RPGP-

CAGCGGCTCCCCGGGGCCCGCCGGGAGGATGCGGACCAGGTCCCCGGTGCGGTAGCGGAT

21841 + + + + + + 21900

GTCGCCGAGGGGCCCCGGGCGGCCCTCCTACGCCTGGTCCAGGGGCCACGCCATCGCCTA
14 LPEGPGAPLIRVLDGTRYRI-

CAGTGGTTTGATGCCGTCCACCAGCATGGTGAGGACGAGTTCGCCCTCTCCCGTGTCGCC

21901 + + + + + + 21960

GTCACCAAACTACGGCAGGTGGTCGTACCACTCCTGCTCAAGCGGGAGAGGGCACAGCGG
14 LPKIGDVLMTLVLEGEGTDG-

GACCACGGCGCCGGTGTCCGGTTCGACGAGTTCGGTCAAGTAGTTGGGCTGGGCGAGGTG

21961 + + + + + + 22020

CTGGTGCCGCGGCCACAGGCCAAGCTGCTCAAGCCAGTTCATCAACCCGACCCGCTCCAC
14 VVAGTDPEVLETLYNPQALH-

GAGCGCTCCGGTGTCCGCTCCGGTGGCGATGCACAGGGCTTCCTGGGAGCCGTAGAGCGT

22021 + + + + + + 22080

CTCGCGAGGCCACAGGCGAGGCCACCGCTACGTGTCCCGAAGGACCCTCGGCATCTCGCA
14 LAGTDAGTAI CLAEQSGYLT-

GGGCCGCACGACGGCTTGCGGCCAGAGGGTCGCCACGTTGTCGGCGAACTGCGGGGTGCA

22081 + + + + + + 22140

CCCGGCGTGCTGCCGAACGCCGGTCTCCCAGCGGTGCAACAGCCGCTTGACGCCCCACGT
14 PRVVAQPWLTAVNDAFQ PTC-

GATCTCACCCAGCGTGAGGAAGAGCTTCACGGGAAGCCGGGC CAGGT CGTAGCCGTAGTG

22141 + + + + + + 22200

CTAGAGTGGGTCGCACTCCTTCTCGAAGTGCCCTTCGGCCCGGTCCAGCATCGGCATCAC
14 IEGLTLFLKVPLRALDYGYH-

CAGGGCCGCCTTGGCAAGGCTCAGGCACAGCGCCGGAGCACAGACGACGACCTCGACCTC

22201 + + + + + + 22260

GTCCCGGCGGAACCGTTCCGAGTCCGTGTCGCGGCCTCGTGTCTGCTGCTGGAGCTGGAG
14 LAAKALS LCLAPACVVVEVE-

CAGCTCCTCGATCAGCCGCAGCGCCTTACGGAATCCCACCCTGGGGGACTCGGGCCAGAT

22261 + + + + + + 22320

GTCGAGGAGCTAGTCGGCGTCGCGGAATGCCTTAGGGTGGGACCCCCTGAGCCCGGTCTA
14 LEEILRLAKRFGVRPSEPWI-

28

CTTGACGTGACAGGCCCCCAGCTCCGCTGCCACCGCGGTGAACACGTCCCCGAACGCGTA

22321 + + + + + + 22380

GAACTGCACTGTCCGGGGGTCGAGGCGACGGTGGCGCCACTTGTGCAGGGGCTTGCGCAT
14 KVH CAGL EAAVAT FVDG F A Y -

CAGCTCCGACGGCCCCATCAGGCCCACGACGGGCATCCGCCCCCCGAACCTCGCTTCCAG

22381 + + + + + + 22440

GTCGAGGCTGCCGGGGTAGTCCGGGTGCTGCCCGTAGGCGGGGGGCTTGGAGCGAAGGTC
14 LESPGMLGVVPMRGGFRAEL-

CATGCGGCGCCAGGACTCCCGGACGGCGATGTTGCTGGTCGCGATGTCCTTCTCGCCGCG

22441 + + + + + + 22500

GTACGCCGCGGTCCTGAGGGCCTGCCGCTACAACGACCAGCGCTACAGGAAGAGCGGCGC
14 MRRWSERVAINSTAIDKEGR-

TGGGCACGGGGTGGCCGCCCCGGTGGTCCCGGTGGTCTCGTAGTAGATGCGTGCTTCGTG

22501 + + + + + + 22560

ACC CGTGC CCCAC CGGCGGGGCCACCAGGGC CAC CAGAGCATCATCTACGCACGAAGCAC
14 PCPTAAGTTGTTEYY I R A E H -

CAGCGGGCCCGACAGGACGTCGTGCATCTCCCGCCGCAGGTCGTCCTTGGTGGTGAAGGG

22561 + + + + + + 22620

GTCGCCCGGGCTGTCCTGCAGCACGTAGAGGGCGGCGTCCAGCAGGAACCACCACTTCCC
14 LPGSLVDHMERRLDDKTTFP-

CAGGTCCGCCAGGTTCGCGGGGGTGACGGCCTCGACGTCCACGCCTGCCAGATGGCGGCG

22621 + + + + + + 22680

GTCCAGGCGGTCCAAGCGCCCCCACTGCCGGAGCTGCAGGTGCGGACGGTCTACCGCCGC
14 LDALNAPTVAEVDVGALHRR-

GTAGAACGGCGAGCGGCGGGTGACGTGGCGCAGTACGGCCGTCAGCCGTTCGCCCTCCCA

22681 + + + + + + 22740

CATCTTGCCGCTCGCCGCCCACTGCACCGCGTCATGCCGGCAGTCGGCAAGCGGGAGGGT
14 YFPS RRTVHRLVATLREGEW-

GCGCTCGCGGTCGGCGGCGGTGAGTTCGCCGCGGTAGAACGCGTCGCTCACCTGCCCGTA

22741 + + + + + + 22800

CGCGAGCGCCAGCCGCCGCCACTCAAGCGGCGCCATCTTGCGCAGCGAGTGGACGGGCAT
14 RERDAATLEGRYFAD SVQGY-

GGCGGACCAGAACTCGCTGTCCGCGTCGGGGTCCAGCGGCCCGGTCCCGCCGGGACCGGG

22801 + + + + + + 22860

CCGCCTGGTCTTGAGCGACAGGCGCAGCCCCAGGTCGCCGGGCCAGGGCGGCCCTGGCCC
14 ASWFESDADPDLPGTGGPGP-

CCGCCGGCCGTCTCTCACGGCTGTGCCTGGAGTTCGTTGAGCGCGAGGCCGACCCGCTCG

22861 + + + + + + 22920

GGCGGCCGGCAGAGAGTGCCGACACGGACCTCAAGCAACTCGCGCTCCGGCTGGGCGAGC

14 -< RRGDRM-

21-* * PQAQLENLALGVRE -

TTGACCTCGTTGGAGGCCAGCACGTCCGAACGGCCGGTGAGCCGACGGTGTTCGTCGAGC

22921 + + + + + + 22980

AACTGGAGCAACCTCCGGTCGTGCAGGCTTGCCGGCCACTCGGCTGCCACAAGCAGCTCG
21 NVENSALVDSRGTLRRHEDL

AGTTCGATCATGTCCGTCATCCTCTCGACCAGGCGCGAGACGTTGGTGAGGCCCTCCTCG

22981 + + + + + + 23040

TCAAGCTAGTACAGGCAGTAGGAGAGCTGGTCCGCGCTCTGCAACCACTCCGGGAGGAGC
21 LEIMDTMREVLRSVNTLGEE

TCCTTGAGCGCGTCGCCCCGGTGCAGCGCGTGCACCGTCGCCGGGAAGCCGCTGCCCACC

23041 + + + + + + 23100

AGGAACTCGCGCAGCGGGGCCACGTCGCGCACGTGGCAGCGGCCCTTCGGCGACGGGTGG
21 DKLADGRHLAHVTAPFGSGV

29

AGGATCATCCGGTTGAGCAGGGCATTGACGGTCAGCTGAGCCCATACCTCGCCGGCGCTG

23101 + + + + + + 23160

TCCTAGTAGGCCAACTCGTCCCGTAACTGCCAGTCGACTCGGGTATGGAGCGGCCGCGAC
21 LIMRNLLANVTLQAWVEGAS-

TAGCGGCGGGCGACCGAGATGATCCCCGCGACCTTGTTGCTCAGCGGCCGGTCGAAGCGC

23i5i + + + + + + 23220

ATCGCCGCCCGCTGGCTCTACTAGGGGCGCTGGAACAACGAGTCGCCGGCCAGCTTCGCG
21 YRRAVS I IGAVKNSLPRDFR-

AGATAACCGACTCCGGCACGCTCGATGAAGGTCTGCATGAGGCTGGCCGTGCCGAATCCG

23221 i — — i 1 1 1 h 23280

TCTATTGGCTGAGGCCGTGCGAGCTACTTCCAGACGTACTCCGACCGGCACGGCTTAGGC
21 LYGVGARE I FTQMLSATGFG

TGCACGGGCGCCGCGAAGATGATCCCGTCCGCCGCGACCATCTTCGCCACGACCTCGGGC

2328I + + + + + + 23340

ACGTGCCCGCGGCGCTTCTACTAGGGCAGGCGGCGCTGGTAGAAGCGGTGCTGGAGCCCG
21 HVPAAFI IGDAAVMKAVVEP

ACCCCGTCGGCCAGGGTGCAGGCCACCGGCCTGTCGTTGCAGTCCCCGCAGGGCCCGCAC

23341 + + + + + + 23400

21 VGDALTCAVPRDNCDGCPGC

CGCTCCATCCTGATCGAGCGCAGGTCGACGGCCTCGAAGTCGACGCCGCGGTTCTCTGCT

23401 + + + + + + 23460

GCGAGGTAGGACTAGCTCGCGTCCAGCTGCCGGAGCTTCAGCTGCGGCGCCAAGAGACGA

21 REMRI SRLDVAEFDVGRNEA-

ACGCGTGCCGCGTGCCGCAGTACGTCGGCGGTGTTGCCGTCACGTTCCGAACCGTTGATC

234 6 i + + + + + + 23520

TGCGCACGGCGCACGGCGTCATGCAGCCGCCACAACGGCAGTGCAAGGCTTGGCAACTAG
21 VRAAHRLVDATNGDRESGNI

GCGAGGATCTTGAGTTGTGCGCTCACGAGGGGCCTCCTTGGTGAGTCAGGTGCGCTCGGC

23521 + + + + + + 23580

CGCTCCTAGAACTCAACACGCGAGTGCTCCCCGGAGGAACCACTCAGTCCACGCGAGCCG
!3_* * T R E A -

21-< ALIKLQASM-

GGTCGGCTCGGGGGAACTGTCTGGCCGCCGCTGGTCCGGGAGCCGCAGGGCCGGCTCGGC

23581 + + + + + + 23640

CCAGCCGAGCCCCCTTGACAGACCGGCGGCGACCAGGCCCTCGGCGTCCCGGCCGAGCCG

13 TPEPSSDPRRQDPLRLAPEA-

GGGGGCGGGAGGAAGACCGCCCCGCGGCGGGCCGCCACGCTCGCCGAACCGGATGAGGGG

23641 + + + + + + 23700

CCCCCGCCCTCCTTCTGGCGGGGCGCCGCCCGGCGGTGCGAGCGGCTTGGCCTACTCCCC

13 PAP PLGGRPPGGREGFRI LP-

CTTCTCGACGAGATAGAAGCTGATGGTCGCCAGCACGACGCTGATCGAGATCGTGAAGAG

23701 i | | 1 1 + 23760

GAAGAGCTGCTCTATCTTCGACTACCAGCGGTCGTGCTGCGACTAGCTCTAGCACTTCTC
13 KEVLYFS ITALVVS I S ITFL-

GAACAGTTCCCAGAACCCCATGTCACCCCGGAATTCCGGCGTTGGCACGGGAGACTTGCC

237 6 i + + + + + + 23820

CTTGTCAAGGGTCTTGGGGTACAGTGGGGCCTTAAGGCCGCAACCGTGCCCTCTGAACGG
13 FLEWFGMDGRFE PTPVPSKG-

GAAGATGCTGCCGTTCCTGAGCCAGAGGTTGATCACGATCTCGTGCCAGAGGTAGACGCC

23821 + + + + + + 23880

CTTCTACGACGGCAAGGACTCGGTCTCCAACTAGTGCTAGAGCACGGTCTCCATCTGCGG
13 FISGNRLWLNIVIEHWLYVG-

GAGGGAGATCTGGCCGAGGAAGAGGATCGGCTTGCTGGTGAAGAGCGCGTCCGAGAACCG

30

23881 + + + + + + 23940

CTCCCTCTAGACCGGCTCCTTCTCCTAGCCGAACGACCACTTCTCGCGCAGGCTCTTGGC
13 LS IQGLFLI PKSTFLADS F R -

GGACTCGGCGCCGGGGACCGTCATCGGTGCCAGGAGCAGCAGGGTGAAGGAGGTCAGGAT

23941 + + + + + + 24000

CCTGAGCCGCGGCCCCTGGCAGTAGCCACGGTCCTCGTCGTCCCACTTCCTCCAGTCCTA
13 S EAGPVTMPALLLLTF STL I -

GAAGTGGTCGACGAGCTCCTGGGCCAGGGCCGCGTTGTCGCCCATGCCCGGGATGCCGAT

24001 + + + + + + 24060

CTTCACCAGCTGCTCGAGGACCCGGTCCCGGCGCAACAGCGGGTACGGGCCCTACGGCTA
13 FHDVLEQALAANDGMGP I G I -

GGGCTTGGTGGCGTAGAGGAGGTACAGCGGGATGAGCGGGACCCAGCAGATCAGCGGGCG

24061 + + + + + + 24120

CCCGAACCACCGCATCTCCTCCATGTCGCCCTACTCGCCCTGGGTCGTCTAGTCGCCCGC
13 PKTAYliLYLP ILPVWCILPR-

CCGGATCACGAAACGGTAGAAGCCCGGGGTCCCTGGCGTCGCCTCGGCGTACGCGGAGTA

24121 + + + + + + 24180

GGCCTAGTGCTTTGCCATCTTCGGGCCCCAGGGACCGCAGCGGAGCCGCATGCGCCTCAT
13 RIVFRYFGPTGPTAEAYASY-

GATGGCCAGTGCCATGCCCGCGGCGAAGCAGCCGGCGTAGTAGGGCGGCCAGTACCACTG

24181 + + + + + + 24240

CTACCGGTCACGGTACGGGCGCCGCTTCGTCGGCCGCATCATCCCGCCGGTCATGGTGAC
13 IALAMGAAFCGAYY P PWYWQ-

CATCGTCGCGCCGGTGGAGGGGAGGTTGGTGTACGTGACCCAGCCGATGGCCATGACTTC

24241 + + + + + + 24300

GTAGCAGCGCGGCCACCTCCCCTCCAACCACATGCACTGGGTCGGCTACCGGTACTGAAG
13 MTAGTS PLNTYTVWG IAMVE-

CAGCGCGGCCAGCGGCAGCAGGAGGCGGCGTGCCTTCTGCCCGGGAGTGCTGCCGCCCCG

24301 + + + + + + 24360

GTCGCGCCGGTCGCCGTCGTCCTCCGCCGCACGGAAGACGGGCCCTCACGACGGCGGGGC
13 LAAL PLLLRRAKQGPT SGGR-

CGCGAGCCGGTGGCCGATCCAGGCGATCAGCGGCAGGGCGAGGTAGAACGTGAACTCGGC

24361 + + + + + + 24420

GCGCTCGGCCACCGGCTAGGTCCGCTAGTCGCCGTCCCGCTCCATCTTGCACTTGAGCCG
13 ALRHGIWAILPLALYFTFEA-

GGGGACCGTCCAGGTGGGCTCGATGCCGTGCATCGGCTGGCCCTCGGGCAGATAGAAGTG

24421 + + + + + + 24480

CCCCTGGCAGGTCCACCCGAGCTACGGCACGTAGCCGACCGGGAGCCCGTCTATCTTCAC
13 PVTWTPE IGHMPQGEPLYFH-

CATGAGCAGCACGGGCCGCAGGACGTCGCTGACGCTGTCGATCTCGAACCAGTTGTAGCC

24481 + + + + + + 24540

GTACTCGTCGTGCCCGGCGTCCTGCAGCGACTGCGACAGCTAGAGCTTGGTCAACATCGG
13 MLLVPRLVDSVSDIEFWNYG-

GGGGATTGCG7^AGACGAGCAACAGGTAGTAGGCGGGCAGGATGCGCAGGGCCCGGCGTTT

24541 + + + + + + 24600

CCCCTAACGCTTCTGCTCGTTGTCCATCATCCGCCCGTCCTACGCGTCCCGGGCCGCAAA
13 P IAFVLLLYYAPLI RLARRK-

GAGGAACCGTCCGGTGGCGGGCCGCTTCGTCCCACTGATGGTGACGCGGGCGTAGGGCTT

24601 + + + + + + 24660

CTCCTTGGCAGGCCACCGCCCGGCGAAGCAGGGTGACTACCACTGCGCCCGCATCCCGAA
13 LFRGTAPRKTGS I TVRAYPK-

GTACAGCATCATTCCGGACAGAGCGAAGAAGGGGGAAGGCATACCCCCAGACCGTCCGCG
24661 + + + + + + 24720

31

CATGTCGTAGTAAGGCCTGTCTCGCTTCTTCCCCCTTCCGTATGGGGGTCTGGCAGGCGC
13 -< YLMMGSLAFFPSPM-

AGGACGCCCCAGAACGGTTTGCCCGGCTCACCGACGAAGCTGCCCACTCCGGCCTGGAAG

24721 + + + + + + 24780

TCCTGCGGGGTCTTGCCAAACGGGCCGAGTGGCTGCTTCGACGGGTGAGGCCGGACCTTC

GCGACGTGGTAGACGACCACACCCAGCGCGAGGACACCTCGCAGTCCCTCGAACTTCGGT

24781 + + + + + + 24840

CGCTGCACCATCTGCTGGTGTGGGTCGCGCTCCTGTGGAGCGTCAGGGAGCTTGAAGCCA

ATTCGCTTGCTTTTTGCGCCACCTGCGTCGCGAAGGACGTCCCCCATGGAACAGTCCCCT

24841 + + + + + + 24900

TAAGCGAACGAAAAACGCGGTGGACGCAGCGCTTCCTGCAGGGGGTACCTTGTCAGGGGA

TTCCCTTGGCACTTGCTCGTTGACTTCCCGAAATAGTCGGGTCTGCGGAGTGTGAGCCGC

24901 + + - + + + + 24960

AAGGGAACCGTGAACGAGCAACTGAAGGGCTTTATCAGCCCAGACGCCTCACACTCGGCG

ATCTCCAATCGTGCTGTTCCGGTGCTCAGGACGACTTGTTTCGGCCTGAGTGGGAAGGCA

24961 + + + + + + 25020

TAGAGGTTAGCACGACAAGGCCACGAGTCCTGCTGAACAAAGCCGGACTCACCCTTCCGT
12-* *SSKNRGSHSP

GCCACCCCCGCCGCCCCGCCTCGGCCAGACCGGGGGCCGAGGAGTCCCGTTCCGAGAGGA

25021 + + + + + + 25080

CGGTGGGGGCGGCGGGGCGGAGCCGGTCTGGCCCCCGGCTCCTCAGGGCAAGGCTCTCCT
12 LWGRRGAEALGPASSDRESL

TCGGAGTGATCTCCGGCGGCCAGGCGATGCCCACCTCCGGATCCAGCGGATTCAAGCCAT

25081 + + + + + + 25140

AGCCTCACTAGAGGCCGCCGGTCCGCTACGGGTGGAGGCCTAGGTCGCCTAAGTTCGGTA
12 IPTIEPPWAIGVEPDLPNLG

GTTCGAGCCGGGGGTCGTAGGCCGCCGAGCACAGGTAGACGATCACCGCCTCGTCGCTCA

25141 + + + + + + 25200

CAAGCTCGGCCCCCAGCATCCGGCGGCTCGTGTCCATCTGCTAGTGGCGGAGCAGCGAGT
12 HELRPDYAASCLYVIVAEDS

GCGTGAGGAATCCGAAGCCCAGCCCCGCGGAGACGTACAGCGCCCGTCCGTTCTCCTCGC

25201 + + + + + + 25260

CGCACTCCTTAGGCTTCGGGTCGGGGCGCCTCTGCATGTCGCGGGCAGGCAAGAGGAGCG
12 LTLFGFGLGASVYLARGNEE

CGAGCTCCACGGTCCGCCAGCCGCCGAAGGTGGGCGACCCCACCCGGATGTCGACCACGG

25261 + + + + + + 25320

GCTCGAGGTGCCAGGCGGTCGGCGGCTTCCACCCGCTGGGGTGGGCCTACAGCTGGTGCC
12 GLEVTRWGGFTPSGVRI DVV

CGCCGAACACGCTGCCGCGCAGGCAGCTGAAGTACTTGGCCTGGCCGGGTACGCCCCCGG

25321 + + + + + + 25380

GCGGCTTGTGCGACGGCGCGTCCGTCGACTTCATGAACCGGACCGGCCCATGCGGGGGCC
12 AGFVSGRLCSFYKAQGPVGG

CGAAGTGGATGCCCCGCAGCACCCCGTGGGAGGAGATCGCGCAGTTCGCCTGCCGCAGGT

25381 + + + + + + 25440

GCTTCACCTACGGGGCGTCGTGGGGCACCCTCCTCTAGCGCGTCAAGCGGACGGCGTCCA
12 AFHIGRLVGHSSIACNAQRL

CGAAGGAGTGGCCTACGGTGCGGCGGAAGGGCTCGCCCTGGAACCACTCGCGAAACGAGC

25441 + + + + + + 25500

GCTTCCTCACCGGATGCCACGCCGCCTTCCCGAGCGGGACCTTGGTGAGCGCTTTGCTCG
12 DFSHGVTRRFPEGQFWERFS

CCCGTTCGTCACGGAAGACCTGCTTCTCCTCCGTCCACGCTCCCGAGATCCCGATCGGCT
25501 + + + + + + 25560

32

GGGCAAGCAGTGCCTTCTGGACGAAGAGGAGGCAGGTGCGAGGGCTCTAGGGCTAGCCGA
GREDRFVQKEETWAGS I GI P

TCATCGCTGGCCCCTTCTCTCGACTTCTCTCGACGACTCGCGGGAGGCGGCCGAGGGGTC

25561 + + + + + + 25620

AGTAGCGACCGGGGAAGAGAGCTGAAGAGAGCTGCTGAGCGCCCTCCGCCGGCTCCCCAG

< K M -

CGCCGGGCCCGTGGGAACGCCGCAGTCTAGATGCGGCGGCACCGGGGGCAGGGGGGTGCG

25621 + + + + + + 25680

GCGGCCCGGGCACCCTTGCGGCGTCAGATCTACGCCGCCGTGGCCCCCGTCCCCCCACGC

GACGACGTCCGCCCCACCTCAGCACACCGGGAGATGCAGGTCGGTGACGGGCGACGTGAC

25681 + + + + + + 25740

CTGCTGCAGGCGGGGTGGAGTCGTGTGGCCCTCTACGTCCAGCCACTGCCCGCTGCACTG

GATGCAACGGTCCGAGGCCCGGTTGCCCGGACGACGGCCCACAGAGCCATCGGAGCAACG

25741 + + + + + + 25800

CTACGTTGCCAGGCTCCGGGCCAACGGGCCTGCTGCCGGGTGTCTCGGTAGCCTCGTTGC

GAGGCGGACCGCAGATGACCAAGCACGCCCGTGACCGCGCGGTAGTCCTCGGCGCAGGGA

25801 + + + + + + 25860

CTCCGCCTGGCGTCTACTGGTTCGTGCGGGCACTGGCGCGCCATCAGGAGCCGCGTCCCT
.> MTKHARDRAVVLGAGM-

TGGCGGGGCTGCTCGCCGCGCGCGTCCTGTCCGAGACGTACAAGGAAGTGCTGGTGATCG

25861 + + + + + + 25920

ACCGCCCCGACGAGCGGCGCGCGCAGGACAGGCTCTGCATGTTCCTTCACGACCACTAGC
AGLLAARVLSETYKEVLVID-

ACCGGGACCGGTTGGGCGGCACGGAGCAGCGCCGCGGTGTCCCGCACGGACGCCACGCCC

25921 + + + + + + 25980

TGGCCCTGGCCAACCCGCCGTGCCTCGTCGCGGCGCCACAGGGCGTGCCTGCGGTGCGGG
RDRLGGTEQRRGVPHGRHAH-

ATGCGCTGCTGGCCAAGGGACAGCAGATCCTCAACGAACTCTTCCCCGGACTCGACACCG

2598I + + + + + + 26040

TACGCGACGACCGGTTCCCTGTCGTCTAGGAGTTGCTTGAGAAGGGGCCTGAGCTGTGGC
ALLAKGQQI LNELFPGLDTE-

AACTCACCTCGGCCGGAATCCCCGCCGGGGACATCGCCGGGAACCTGCGGTGGTACTTCA

26041 + + + + + + 26100

TTGAGTGGAGCCGGCCTTAGGGGCGGCCCCTGTAGCGGCCCTTGGACGCCACCATGAAGT

LTSAGI PAGD IAGNLRWYFN-

ACGGCCGCCGGCTCCAGCCCTTCGACACCGGGCTGATCAGCGTCTCGGCGACGAGGCCCG

26101 + + + + + + 26160

TGCCGGCGGCCGAGGTCGGGAAGCTGTGGCCCGACTAGTCGCAGAGCCGCTGCTCCGGGC
GRRLQPFDTGLI SVSATRPE-

AGCTGGAGTCCCACGTGCGCGCACGGGTCGCCGCGCTGCCACAGGTGAAGATCATGGACG

26161 + + + + + + 26220

TCGACCTCAGGGTGCACGCGCGTGCCCAGCGGCGCGACGGTGTCCACTTCTAGTACCTGC
L E SHVRARVAAL PQVKI MDG-

GGTGCGTGATCCGGGGCCTGACCGCCTCGGCCGACCGCAGCCGCGTCACCGGTGTCGAGG

26221 j | — | -\ — — i — — h 26280

CCACGCACTAGGCCCCGGACTGGCGGAGCCGGCTGGCGTCGGCGCAGTGGCCACAGCTCC
CV I RGLTASADRS RVTGVEV-

TGGTCGACGAGTCGGGTACGGACACCCCGACGCGCCTGGAGGCCGACCTCGTCGTCGACG

26281 + + + + + + 26340

ACCAGCTGCTCAGCCCATGCCTGTGGGGCTGCGCGGACCTCCGGCTGGAGCAGCAGCTGC

VDESGTDTPTRLEADLVVDV-
TCACGGGGCGCGGCTCGCGGACTCCCGCCTGGCTGGAGGAGTTCGGATACGAGCGGCCCG

33

26341 + + + + + + 26400

AGTGCCCCGCGCCGAGCGCCTGAGGGCGGACCGACCTCCTCAAGCCTATGCTCGCCGGGC
20 TGRGSRTPAWLEEFGYERPA-

CGGAGGACCGCTTCAAGATCGATCTGGCGTACACCACGCGCCACTTCAAGCTCAAGGAAG

26401 + + + + + + 26460

GCCTCCTGGCGAAGTTCTAGCTAGACCGCATGTGGTGCGCGGTGAAGTTCGAGTTCCTTC
20 EDRFKIDLAYTTRHFKLKED-

ACCCCTACGGCACGGACCTGTCGATCAACCCGGTGGCATCGCCGAGCAACCCGCGCGGCG

26461 + + + + + + 26520

TGGGGATGCCGTGCCTGGACAGCTAGTTGGGCCACCGTAGCGGCTCGTTGGGCGCGCCGC
20 PYGTDLS I NPVAS PSNPRGA-

CGTTCTTCCCCCGGCTCGCGGACGGCAGCTCCCAGCTCTCCCTCACCGGAATCCTCGGCG

26521 + + + + + + 26580

GCAAGAAGGGGGCCGAGCGCCTGCCGTCGAGGGTCGAGAGGGAGTGGCCTTAGGAGCCGC
20 FFPRLADGS SQLSLTG I LGD-

ACCACCCGCCCACCGACGACGAGGGCTTCCTGGCGTTCGCCAAGTCGCTTGCCGCGCCGG

26581 + + + + + + 26640

TGGTGGGCGGGTGGCTGCTGCTCCCGAAGGACCGCAAGCGGTTCAGCGAACGGCGCGGCC
20 HP PTDDEGFLAFAKS LAAPE-

AGATCTACCGGGCCGTCCGCGATGCCGAACCTCTCGACGAACCGGTCACCTTCCGCTTCC

26641 + + + + + + 26700

TCTAGATGGCCCGGCAGGCGCTACGGCTTGGAGAGCTGCTTGGCCAGTGGAAGGCGAAGG
20 I YRAVRDAEPLDEPVTFRFP-

CGGCGAGCGTCCGCCGCCGTTACGAGAGGCTGCGCCGTTTCCCCGGCGGGTTCCTCGTCA

26701 + + + + + + 26760

GCCGCTCGCAGGCGGCGGCAATGCTCTCCGACGCGGCAAAGGGGCCGCCCAAGGAGCAGT
20 ASVRRRYERLRRFPGGFLVM-

TGGGCGACGGCGTGTGCAGCTTCAACCCCGTCTACGGCCAGGGCATGACGGTCGCCGCCC

26761 + + + + + + 26820

ACCCGCTGCCGCACACGTCGAAGTTGGGGCAGATGCCGGTCCCGTACTGCCAGCGGCGGG
20 GDGVC S FNPVYGQGMTVAAL-

TGGAGGCCGTGGCGCTGCGGGACCACTTGCGCGACGCCCCGGACCCCGACGCCCTGCGCT

26821 + + + + + + 26880

ACCTCCGGCACCGCGACGCCCTGGTGAACGCGCTGCGGGGCCTGGGGCTGCGGGACGCGA
20 EAVALRDHLRDAPDPDALRF-

TCTTCCGGCGTATCTCCACGGTCATCGACGTTCCGTGGGACATCGCCGCCGGAGCGGATC

26881 + + + + + + 26940

AGAAGGCCGCATAGAGGTGCCAGTAGCTGCAAGGCACCCTGTAGCGGCGGCCTCGCCTAG
20 FRRI STVIDVPWDIAAGADL-

TGAACTTCCCCGGGGTGGAGGGCCCCCGCACCATGAAGGTGAAGATGGCCAACGCCTACA

26941 + + + + + + 27000

ACTTGAAGGGGCCCCACCTCCCGGGGGCGTGGTACTTCCACTTCTACCGGTTGCGGATGT
20 NF PGVEG PRTMKVKMANAYM-

TGGCCCGCCTGCACGCAGCGGCAGCCGTCGACGGCGCGGTGACCGGGGCGTTCTTCCGGG

27001 + + + + + + 27060

ACCGGGCGGACGTGCGTCGCCGTCGGCAGCTGCCGCGCCACTGGCCCCGCAAGAAGGCCC
20 ARLHAAAAVDGAVTGAF FRV-

TGGCCGGGCTGGTGGACCCCCCGCAGGCCCTGATGCGCCCCTCCCTCGCCCTGCGGGTCA

27061 + + + - - + + + 27120

ACCGGCCCGACCACCTGGGGGGCGTCCGGGACTACGCGGGGAGGGAGCGGGACGCCCAGT
20 AGLVD P PQALMR P S LAL RVM-

TGCGCAACTCCTCGGCGAAGCCGTCGGTCCCTTCGGGCGCCGCCGTATGACCGCGCGGCC
27121 + + + + + + 27180

34

ACGCGTTGAGGAGCCGCTTCGGCAGCCAGGGAAGCCCGCGGCGGCATACTGGCGCGCCGG
20-* RNS SAKPSVPSGAAV*-

CGTCCGGGGCGGCTGCCGGGGCCAGGAGCCGACATGCGGGTGATGATCACGGTGTTCCCG

27181 + + + + + + 27240

GCAGGCCCCGCCGACGGCCCCGGTCCTCGGCTGTACGCCCACTACTAGTGCCACAAGGGC
19 MRVMITVFP

GCGCGGGCGCACTTCCTGCCGCTGGTGCCCTATGCCTGGGCCCTGCAGAGCGCGGGCCAC

27241 + + + + + + 27300

CGCGCCCGCGTGAAGGACGGCGACCACGGGATACGGACCCGGGACGTCTCGCGCCCGGTG
19 ARAHFLPLVPYAWALQSAGH

GAGGTATGTGTCGTGGCGCCCCCGGGCTATCCCACCGGGGTGGCCGACCCCGACTTCCAC

27301 + + + + + + 27360

CTCCATACACAGCACCGCGGGGGCCCGATAGGGTGGCCCCACCGGCTGGGGCTGAAGGTG
19 EVCVVAP PGYPTGVADPDFH

GAGGCCGTCACCGCGGCCGGCCTGAAGTCGGTGACCTGCGGGCAGCCGCAGCCGCTGGCG

27361 + + + + + + 27420

CTCCGGCAGTGGCGCCGGCCGGACTTCAGCCACTGGACGCCCGTCGGCGTCGGCGACCGC
19 EAVTAAGLKSVTCGQPQPLA

GTCCACGACCGCGACGACCCCGGCTACGCGGCGATGCTGCCGACCGCGGCGGAGTCGGAG

27421 + + + + + + 27480

CAGGTGCTGGCGCTGCTGGGGCCGATGCGCCGCTACGACGGCTGGCGCCGCCTCAGCCTC
19 VHDRDDPGYAAMLPTAAESE

CGCTACGTGGCGGCCCTCGGGATCAGCGAGAAGGAGCGCCCCACCTGGGACGTCTTCTAC

27481 + + + + + + 27540

GCGATGCACCGCCGGGAGCCCTAGTCGCTCTTCCTCGCGGGGTGGACCCTGCAGAAGATG
19 RYVAALGISEKERPTWDVFY

CACTTCACCTTGCTGGCGATCCGCGACTACCATCCGCCGCGGCCGCGGCAGGACGTGGAC

27541 + + + + + + 27600

GTGAAGTGGAACGACCGCTAGGCGCTGATGGTAGGCGGCGCCGGCGCCGTCCTGCACCTG
19 HFTLLAIRDYHPPRPRQDVD

CAGGTGATCGAGTTCGCCCGGATCTGGCAGCCCGATCTGGTGCTGTGGGACGCCTGGTTC

27601 + + + + + + 27660

GTCCACTAGCTCAAGCGGGCCTAGACCGTCGGGCTAGACCACGACACCCTGCGGACCAAG
19 QVIEFARIWQPDLVLWDAWF

CCCTCGGGCGCGATCGCGGCGCGGGTCAGCGGCGCCGCGCACGCGCGGGTGCTCGTAGCC

27661 + + + + + + 27720

GGGAGCCCGCGCTAGCGCCGCGCCCAGTCGCCGCGGCGCGTGCGCGCCCACGAGCATCGG
19 PSGAIAARVSGAAHARVLVA

CCCGACTACACCGGCTGGGTCACCGAGCGGTTCGCCGCCGCGGGCCCCGCGGCGGGGGCC

27721 + + + + + + 27780

GGGCTGATGTGGCCGACCCAGTGGCTCGCCAAGCGGCGGCGCCCGGGGCGCCGCCCCCGG
19 PDYTGWVTERFAAAGPAAGA

GACCTCCTGGCCGAGACGATGCGGCCGCTGGCCGAGCGGTACGGCGTGGAGGTCGACGAC

27781 + + + + + + 27840

CTGGAGGACCGGCTCTGCTACGCCGGCGACCGGCTCGCCATGCCGCACCTCCAGCTGCTG
19 DLLAETMRPLAERYGVEVDD

GATCTTCTGCTCGGACAGTGGACGGTCAATCCGTTCCCGGCGCCGATGAACCCGCCGACC

27841 + + + + + + 27900

CTAGAAGACGAGCCTGTCACCTGCCAGTTAGGCAAGGGCCGCGGCTACTTGGGCGGCTGG
19 DLLLGQWTVNPFPAPMNPPT

CGGCTCACGAACGTTCCGGTGCGCTACGTGCCCTACACCGGTGCCAGCGTCATGCCCGCG

27901 + + + + + + 27960

GCCGAGTGCTTGCAAGGCCACGCGATGCACGGGATGTGGCCACGGTCGCAGTACGGGCGC

35

RLTNVPVRYVPYTGASVMPA

TGGCTGTACGCGCGGCCGTCGCGGCCGCGGGTGGCGCTGTCGCTCGGAGTGTCCGCGCGG

2796I + + + + + + 28020

ACCGACATGCGCGCCGGCAGCGCCGGCGCCCACCGCGACAGCGAGCCTCACAGGCGCGCC
WLYARPSRPRVALSLGVSAR

GCGTTCCTCAAGGGTGACTGGGGGCGTACCGCCAAACTGCTGGAAGCGGTCGCGGAGCTG

28021 + + + + + + 28080

CGCAAGGAGTTCCCACTGACCCCCGCATGGCGGTTTGACGACCTTCGCCAGCGCCTCGAC
AFLKGDWGRTAKLLEAVAEL

GACATCGAGGTGATCGCCACGCTCAACGACAACCAACTGGCGGAGAGCGGGCCGCTGCCG

28081 + + + + + + 28140

CTGTAGCTCCACTAGCGGTGCGAGTTGCTGTTGGTTGACCGCCTCTCGCCCGGCGACGGC
DIEVIATLNDNQLAESGPLP

GACAACGTCCACACCCTCGACTACGTACCGCTCGACCAGTTGCTGCCCACCTGCTCGGCC

28141 + + + + + + 28200

CTGTTGCAGGTGTGGGAGCTGATGCATGGCGAGCTGGTCAACGACGGGTGGACGAGCCGG
DNVHTLDYVPLDQLLPTCSA

GTCATCCACCACGGATCGACGGGCACCTTCGCCGCGGCGAGCGCGGCCGGGCTGCCCCAG

28201 + + + + + + 28260

CAGTAGGTGGTGCCTAGCTGCCCGTGGAAGCGGCGCCGCTCGCGCCGGCCGGACGGGGTC
VIHHGSTGTFAAASAAGLPQ

GTGGTCTGCGACACCGACGAGCCCCTCCTGCTCTTCGGCGAGGACACCCCCGACGGCATC

28 261 + + + + + + 28320

CACCAGACGCTGTGGCTGCTCGGGGAGGACGAGAAGCCGCTCCTGTGGGGGCTGCCGTAG
VVCDTDEPLLLFGEDTPDGI

GCGTGGGACTTCACCTGCCAGAAGCAGCTCACCGCGACGCTCACCTCCCGCGTGGTCACC

28321 + + + + + + 28380

CGCACCCTGAAGTGGACGGTCTTCGTCGAGTGGCGCTGCGAGTGGAGGGCGCACCAGTGG
AWDFTCQKQLTATLTSRVVT

GACTACGGGGCGGGGGTGCGCGTCGACCACCAGAAGCAGTCCGCCGGACAGATCCGTGAG

28381 + + + + + + 28440

CTGATGCCCCGCCCCCACGCGCAGCTGGTGGTCTTCGTCAGGCGGCCTGTCTAGGCACTC
DYGAGVRVDHQKQSAGQ I RE

CAACTACGCAGGGTGCTCACCGAACCTTCCTTCCGCGAGGGCGCTCGACGGATCCGGGAA

28441 + + + + + + 28500

GTTGATGCGTCCCACGAGTGGCTTGGAAGGAAGGCGCTCCCGCGAGCTGCCTAGGCCCTT
QLRRVLTEPSFREGARRIRE

GACCGGAATTCCGCCCCCAGCCCGGTCGAACTCGTATCGCTCCTGGTAGAACTGACGAAG

28501 + + + + + + 28560

CTGGCCTTAAGGCGGGGGTCGGGCCAGCTTGAGCATAGCGAGGACCATCTTGACTGCTTC
DRNSAPSPVELVSLLVELTK

CGTCATCGCCGTGACAAGGAGGCGGACCGATGAGGATGCTGGTGACGGGCGGAGCGGGTT

28561 + + + + + + 28620

GCAGTAGCGGCACTGTTCCTCCGCCTGGCTACTCCTACGACCACTGCCCGCCTCGCCCAA

-* RHRRDKEADR*-

> MRMLVTGGAGF-

TCATCGGCTCGCAGTTCGTGCGGGCCACACTGCACGGCGAGCTGCCGGGTTCCGAGGACG

2 8 621 + + + + + + 28680

AGTAGCCGAGCGTCAAGCACGCCCGGTGTGACGTGCCGCTCGACGGCCCAAGGCTCCTGC
IGSQFVRATLHGELPGSEDA-

CCCGGGTGACGGTCCTGGACAAGCTGACGTACTCCGGCAATCCGGCCAACCTCACCTCCG

28681 + + + + + + 28740

GGGCCCACTGCCAGGACCTGTTCGACTGCATGAGGCCGTTAGGCCGGTTGGAGTGGAGGC

36

1 RVTVLDKLTYSGNPANLTSV-

TCGCGGCCCATCCGCGGTACACCTTCGTCCAGGGCGACACCGTCGACCCGCGCGTCGTCG

28741 + + + + + + 28800

AGCGCCGGGTAGGCGCCATGTGGAAGCAGGTCCCGCTGTGGCAGCTGGGCGCGCAGCAGC
1 AAH PRYTFVQGDTVD P RVVD-

ACGAGGTGGTCGCCGGCCACGACGTCATCGTCCACTTCGCGGCGGAGTCGCACGTGGACC

28801 + + + + + + 28860

TGCTCCACCAGCGGCCGGTGCTGCAGTAGCAGGTGAAGCGCCGCCTCAGCGTGCACCTGG
1 EVVAGHDVIVHFAAES HVDR-

GCTCGATCGACACCGCCACCCGGTTCGTCACGACCAACGTGCTCGGGACCCAGACGCTGC

28861 + + + + + + 28920

CGAGCTAGCTGTGGCGGTGGGCCAAGCAGTGCTGGTTGCACGAGCCCTGGGTCTGCGACG
1 SIDTATRFVTTNVLGTQTLL-

TGGAAGCGGCTCTCCGGCACGGGGTCGGCCGGTTCGTGCACGTGTCGACCGACGAGGTCT

28921 + + + + + + 28980

ACCTTCGCCGAGAGGCCGTGCCCCAGCCGGCCAAGCACGTGCACAGCTGGCTGCTCCAGA
1 EAALRHGVGRFVHVS TDEVY-

ACGGGTCGATCGCCTCCGGCTCATGGACCGAGGACACCCCGCTCGCCCCCAACGTCCCCT

28981 + + + + + + 29040

TGCCCAGCTAGCGGAGGCCGAGTACCTGGCTCCTGTGGGGCGAGCGGGGGTTGCAGGGGA
1 GSIASGSWTEDTPLAPNVPY-

ACGCGGCGTCGAAGGCGGGTTCGGACCTGATGGCGCTCGCCTGGCACCGCACCCGGGGCC

29041 + + + + + + 29100

TGCGCCGCAGCTTCCGCCCAAGCCTGGACTACCGCGAGCGGACCGTGGCGTGGGCCCCGG
1 AASKAGSDLMALAWHRTRGL-

TGGACGTCGTCGTCACCCGGTGCACCAACAACTACGGTCCCTACCAGTACCCCGAGAAGG

29101 + + + + + + 29160

ACCTGCAGCAGCAGTGGGCCACGTGGTTGTTGATGCCAGGGATGGTCATGGGGCTCTTCC
1 DVVVTRCTNNYGPYQYPEKV-

TGATCCCGCTCTTCGTCACCAACATCCTCGACGGCTTGCGGGTGCCCCTGTACGGGGACG

29161 + + + + + + 29220

ACTAGGGCGAGAAGCAGTGGTTGTAGGAGCTGCCGAACGCCCACGGGGACATGCCCCTGC
1 IPLFVTNILDGLRVPLYGDG-

GCGCCCACCGCCGGGACTGGCTGCACGTGTCCGACCACTGCCGGGCCATCCAGATGGTCA

29221 + + + + + + 29280

CGCGGGTGGCGGCCCTGACCGACGTGCACAGGCTGGTGACGGCCCGGTAGGTCTACCAGT
1 AHRRDWLHVSDHCRAI QMVM-

TGAACTCCGGCCGGGCCGGGGAGGTCTACCACATCGGCGGCGGCACCGAACTCTCCAACG

29281 + + + + + + 29340

ACTTGAGGCCGGCCCGGCCCCTCCAGATGGTGTAGCCGCCGCCGTGGCTTGAGAGGTTGC
1 NSGRAGEVYHIGGGTELSNE-

AGGAACTCACCGGCCTGTTGCTCACGGCGTGCGGCACCGACTGGTCCTGCGTGGACCGGG

29341 + + + + + + 29400

TCCTTGAGTGGCCGGACAACGAGTGCCGCACGCCGTGGCTGACCAGGACGCACCTGGCCC
1 ELTGLLLTACGTDWS CVDRV-

TGGCCGACCGGCAGGGGCACGACCGCCGCTACTCGCTCGACATCACGAAGATCCGGCAGG

29401 + + + + + + 29460

ACCGGCTGGCCGTCCCCGTGCTGGCGGCGATGAGCGAGCTGTAGTGCTTCTAGGCCGTCC
1 ADRQGHDRRYSLDITKI RQE-

AACTGGGCTACGAGCCCCTGGTCGCCTTCGAGGACGGCCTGGCCGCGACGGTGAAGTGGT

29461 + + + + + + 29520

TTGACCCGATGCTCGGGGACCAGCGGAAGCTCCTGCCGGACCGGCGCTGCCACTTCACCA
1 LGYEPLVAFEDGLAATVKWY-

37

ACCACGAGAACCGTTCGTGGTGGCAGCCGCTGAAGGAAGCGGCCGGCCTCCTGGACGCCG

29521 + + + + + + 29580

TGGTGCTCTTGGCAAGCACCACCGTCGGCGACTTCCTTCGCCGGCCGGAGGACCTGCGGC

1 HENRSWWQPLKEAAGLLDAV-

TCGGCTGACGGCAGCCACCGCTAGGAACACCCCAGGAAAGGAGCCACCTCCGTGACAGCA

29581 + + + + + + 29640

AGCCGACTGCCGTCGGTGGCGATCCTTGTGGGGTCCTTTCCTCGGTGGAGGCACTGTCGT
2-> M T A

1-* G * -

GTCAAGGAGCCGACGTCCCGCGCAGGACGGCGGGAGTGGATCGCTCTCGTCGTCCTCTCC

29641 + + + + + + 29700

CAGTTCCTCGGCTGCAGGGCGCGTCCTGCCGCCCTCACCTAGCGAGAGCAGCAGGAGAGG

2 VKEPTSRAGRREWIALVVLS

TTGCCCACGATGCTGTTGATGCTGGACATCAACGTCCTCATGCTGGCCTTGCCGCAGTTG

29701 + + + + + + 29760

AACGGGTGCTACGACAACTACGACCTGTAGTTGCAGGAGTACGACCGGAACGGCGTCAAC
2 LPTMLLMLDINVLMLALPQL

AGCGAGGATCTCGGCGCGAGCAGCACGCAACAGCTGTGGATCACCGACATCTACGGATTC

29761 + + + + + + 29820

TCGCTCCTAGAGCCGCGCTCGTCGTGCGTTGTCGACACCTAGTGGCTGTAGATGCCTAAG
2 SEDLGASSTQQLWITDIYGF

GCGATCGCCGGCTTCCTGGTGACCATGGGCACCCTCGGCGACCGGATCGGCCGCCGCAGG

29821 + + + + + + 29880

CGCTAGCGGCCGAAGGACCACTGGTACCCGTGGGAGCCGCTGGCCTAGCCGGCGGCGTCC
2 AIAGFLVTMGTLGDRIGRRR

CTCCTGCTCGGGGGCGCGGCCGTCTTCGCGGTCGTGTCCGTCGTCGCCGCGTTCTCCGAC

29881 + + + + + + 29940

GAGGACGAGCCCCCGCGCCGGCAGAAGCGCCAGCACAGGCAGCAGCGGCGCAAGAGGCTG

2 LLLGGAAVFAVVSVVAAFSD

AGCGCGGCGATGCTCGTCGTCAGCCGCGCCGTGCTCGGCGTCGCCGGGGCCACGGTGATG

29941 + + + + + + 30000

TCGCGCCGCTACGAGCAGCAGTCGGCGCGGCACGAGCCGCAGCGGCCCCGGTGCCACTAC
2 SAAMLVVSRAVLGVAGATVM

CCCTCGACGCTCGCGCTCATCAGCAACATGTTCGAGGACCCCAAGGAGCGGGGCACCGCC

30001 + + + + + + 30060

GGGAGCTGCGAGCGCGAGTAGTCGTTGTACAAGCTCCTGGGGTTCCTCGCCCCGTGGCGG
2 PSTLALI SNMFEDPKERGTA

ATCGCCATGTGGGCGAGCGCCATGATGGCCGGAGTCGCCCTCGGGCCCGCCGTCGGCGGC

30061 + + + + + + 30120

TAGCGGTACACCCGCTCGCGGTACTACCGGCCTCAGCGGGAGCCCGGGCGGCAGCCGCCG
2 IAMWASAMMAGVALGPAVGG

CTGGTCCTCGCCGCGTTCTGGTGGGGATCGGTGTTCCTCATCGCCGTTCCGGTGATGCTG

30121 + + + + + + 30180

GACCAGGAGCGGCGCAAGACCACCCCTAGCCACAAGGAGTAGCGGCAAGGCCACTACGAC

2 LVLAAFWWGSVFL IAVPVML

CTGGTGGTGGTCACCGGCCCCGTGCTGCTCACCGAGTCCCGCGACCCGGACGCCGGACGG

30181 + + + + + + 30240

GACCACCACCAGTGGCCGGGGCACGACGAGTGGCTCAGGGCGCTGGGCCTGCGGCCTGCC
2 LVVVTGPVLLTESRDPDAGR

CTGGACCTGCTGAGCGCGGGGCTCTCCCTCGCGACCGTGCTGCCGGTGATCTACGGACTG

30241 + + + + + + 30300

GACCTGGACGACTCGCGCCCCGAGAGGGAGCGCTGGCACGACGGCCACTAGATGCCTGAC
2 LDLLSAGLSLATVLPVIYGL

38

AAGGAGCTGGCCCGGACCGGGTGGGACCCGCTCGCCGCCGGCGCGGTGGTCCTCGGCGTG

30301 + + + + + + 30360

TTCCTCGACCGGGCCTGGCCCACCCTGGGCGAGCGGCGGCCGCGCCACCAGGAGCCGCAC
KELARTGWDPLAAGAVVLGV

ATCTTCGGCGCGCTGTTCGTCCAGCGCCAGCGGCGGTTGGCCGACCCCATGCTGGACCTC

30361 + + + + + + 30420

TAGAAGCCGCGCGACAAGCAGGTCGCGGTCGCCGCCAACCGGCTGGGGTACGACCTGGAG
I FGALFVQRQRRLAD PMLDL

GGCCTCTTCGCCGACCGCACCCTGCGGGCGGGTCTGACGGTCAGTCTGGTCAACGCCGTC

30421 + + + + + + 30480

CCGGAGAAGCGGCTGGCGTGGGACGCCCGCCCAGACTGCCAGTCAGACCAGTTGCGGCAG
GLFADRTLRAGLTVSLVNAV

ATCATGGGCGGGACCGGACTGATGGTCGCCCTGTACCTCCAGACGATCGCCGGTCACTCC

30481 + + + + + + 30540

TAGTACCCGCCCTGGCCTGACTACCAGCGGGACATGGAGGTCTGCTAGCGGCCAGTGAGG
IMGGTGLMVALYLQTIAGHS

CCGTTGGCCGCCGGGCTGTGGCTGCTGATCCCGGCCTGCATGCTCGTCGTGGGCGTACAG

30541 + + + + + + 30600

GGCAACCGGCGGCCCGACACCGACGACTAGGGCCGGACGTACGAGCAGCACCCGCATGTC
PLAAGLWLLI PACMLVVGVQ

CTGTCGAACCTGCTGGCCCAGCGGATGCCCCCTTCCCGGGTGCTGCTGGGGGGACTGCTG

30601 + + + + + + 30660

GACAGCTTGGACGACCGGGTCGCCTACGGGGGAAGGGCCCACGACGACCCCCCTGACGAC
LSNLLAQRMPPSRVLLGGLL

ATCGCGGCCGTCGGACAGCTCCTGATCACCCAGGTGGACACCGAGGACACCGCCCTCCTC

30661 + + + + + + 30720

TAGCGCCGGCAGCCTGTCGAGGACTAGTGGGTCCACCTGTGGCTCCTGTGGCGGGAGGAG
IAAVGQLLITQVDTEDTALL

ATCGCGGCCACCACCCTGATCTACTTCGGCGCCTCACCGGTGGGGCCGATCACCACGGGC

30721 + + + + + + 30780

TAGCGCCGGTGGTGGGACTAGATGAAGCCGCGGAGTGGCCACCCCGGCTAGTGGTGCCCG
IAATTLIYFGASPVGPITTG

GCGATCATGGGAGCCGCGCCCCCGGAGAAGGCGGGTGCCGCCTCGTCGCTGTCCGCCACC

30781 + + + + + + 30840

CGCTAGTACCCTCGGCGCGGGGGCCTCTTCCGCCCACGGCGGAGCAGCGACAGGCGGTGG
AIMGAAPPEKAGAASSLSAT

GGCGGCGAGTTCGGAGTGGCGCTCGGCATCGCGGGCCTGGGGAGTCTGGGCACCGTCGTG

30841 + + + + + + 30900

CCGCCGCTCAAGCCTCACCGCGAGCCGTAGCGCCCGGACCCCTCAGACCCGTGGCAGCAC
GGEFGVALGIAGLGSLGTVV

TACAGCGCCGGGGTCGAGGTGCCGGACGCGGCCGGGCCCGCCGACGCCGACGCCGCGCAG

30901 + + + + + + 30960

ATGTCGCGGCCCCAGCTCCACGGCCTGCGCCGGCCCGGGCGGCTGCGGCTGCGGCGCGTC
YSAGVEVPDAAGPADADAAQ

GAGAGCATCGCCGGCGCCCTGCACACGGCCGGTCAGCTGGCACCGGGCAGCGCCGACGCC

30961 + + + + + + 31020

CTCTCGTAGCGGCCGCGGGACGTGTGCCGGCCAGTCGACCGTGGCCCGTCGCGGCTGCGG
ESIAGALHTAGQLAPGSADA

CTGCTGGACTCCGCGCGCGCGGCCTTCACCAGCGGCGTGCAGTCCGTCGCCGCCGTCTGC

31021 + + + + + + 31080

GACGACCTGAGGCGCGCGCGCCGGAAGTGGTCGCCGCACGTCAGGCAGCGGCGGCAGACG
LLDSARAAFTSGVQSVAAVC

39

GCCGTGTTCTCCCTGGCGCTCGCCGTCCTCATCGGCACCCGGCTGCGGGACATTTCCGCG

31081 + + + + + + 31140

CGGCACAAGAGGGACCGCGAGCGGCAGGAGTAGCCGTGGGCCGACGCCCTGTAAAGGCGC

2 AVFSLALAVLIGTRLRD I SA

ATGGACCACGGGCACGGCGAGGAACCGGCCGAGAACGACGCTCAACCGGCCACATGAGCG

31141 + + + + + + 31200

TACCTGGTGCCCGTGCCGCTCCTTGGCCGGCTCTTGCTGCGAGTTGGCCGGTGTACTCGC

2- * MDHGHGEEPAENDAQPAT*-

CACTTCCGGAGATGCAACGGCCGCCGTCGAGGTATGAGGATCACCTTCCGGGGTGCACCT

31201 + + + + + + 31260

GTGAAGGCCTCTACGTTGCCGGCGGCAGCTCCATACTCCTAGTGGAAGGCCCCACGTGGA

GCACGGCAACGGAGGCGTAGTGGAGTACTGGAACAGCACGGCGGAGACCATGC CC CGCCA

31261 + + + + + + 31320

CGTGCCGTTGCCTCCGCATCACCTCATGACCTTGTCGTGCCGCCTCTGGTACGGGGCGGT

3- > MEYWNSTAETMPRQ

GGAACTCGAACAGTGGAAGTGGCGCAGGCTCCAGGCCGCCATGGACCACGCCAGAAGGCT

31321 + + + + + + 31380

CCTTGAGCTTGTCACCTTCACCGCGTCCGAGGTCCGGCGGTACCTGGTGCGGTCTTCCGA

3 ELEQWKWRRLQAAMDHARRL

TTCGCCCTTCTGGCGGGAACGACTCCCCGAGAACATCACCTCCATGGCGGACTACGCGGC

31381 + + + + + + 31440

AAGCGGGAAGACCGCCCTTGCTGAGGGGCTCTTGTAGTGGAGGTACCGCCTGATGCGCCG
3 S PFWRERLPENI TSMADYAA-

GCGGGTGCCTCTCCTGCGCAAGGCCGACCTCCTCGCCGCGGAAGCCGCGTCTCCCCCTTA

31441 + + + + + + 31500

CGCCCACGGAGAGGACGCGTTCCGGCTGGAGGAGCGGCGCCTTCGGCGCAGAGGGGGAAT
3 RVPLLRKADLLAAEAASPPY-

CGGCACCTGGCCCTCGCTGGATCCGGCGCTCGGAGTGCGCCATCACCAGACCAGCGGCAC

31501 + + + + + + 31560

GCCGTGGACCGGGAGCGACCTAGGCCGCGAGCCTCACGCGGTAGTGGTCTGGTCGCCGTG
3 GTWPSLDPALGVRHHQTSGT-

CAGCGGTAACCCCCCCATCCGGACGTTCGACACCGAACGCGACTGGGCCTGGTGCGTGGA

31561 + + + + + + 31620

GTCGCCATTGGGGGGGTAGGCCTGCAAGCTGTGGCTTGCGCTGACCCGGACCACGCACCT
3 SGNPPIRTFDTERDWAWCVD-

CACGTTCTGCACGGCGCTCCACAGCATGGGCGTGCGCCCGCACCACAAGGGTCTGGTGGC

31621 + + + + + + 31680

GTGCAAGACGTGCCGCGAGGTGTCGTACCCGCACGCGGGCGTGGTGTTCCCAGACCACCG
3 TFCTALHSMGVRPHHKGLVA-

GTTCGGCTACGGGCTGTTCGCCGGTTTCTGGGGCATGCACTACGGCCTCGAGCGCATGGG

31681 + + + + + + 31740

CAAGC CGATGCC CGACAAGCGGCCAAAGACCCCGTACGTGATGCCGGAGCTCGCGTAC C C
3 FGYGLFAGFWGMHYGLERMG-

CGCCACGGTCATCCCGGCCGGCGGCCTCGACTCCCGCTCCCGGGTACGGCTGCTGGTCGA

31741 + + + + + + 31800

GCGGTGCCAGTAGGGCCGGCCGCCGGAGCTGAGGGCGAGGGCCCATGCCGACGACCAGCT
3 ATVI PAGGLDSRSRVRLLVD

CTACCAGATCGAGGTGCTCGGCCTCACACCGAGCTATGCGATGCGGCTGATCGAGACGGC

3180 l + + + + + + 31860

GATGGTCTAGCTCCACGAGCCGGAGTGTGGCTCGATACGCTACGCCGACTAGCTCTGCCG
3 YQIEVLGLTPSYAMRLIETA-

CCGCGAGATGGGCATCGACCTCGCCCGCGAGGCTAACGTCCAGATCATCCTGGCCGGGGC
31861 + + + + + + 31920

40

GGCGCTCTACCCGTAGCTGGAGCGGGCGCTCCGATTGCAGGTCTAGTAGGACCGGCCCCG
3 REMGIDLAREANVQI ILAGA-

GGAGCCGCGCTCCGCGTTCACCACCCGCACCATCGAGGAGGCCTTCGGCGCCCGGGTCTT

31921 + + + + + + 31980

CCTCGGCGCGAGGCGCAAGTGGTGGGCGTGGTAGCTCCTCCGGAAGCCGCGGGCCCAGAA
3 EPRSAFTTRTIEEAFGARVF

CAACGCCGCGGGCACCACTGAGTTCGGGGGGGTGTTCATGTTCGAGTGCACCGCCCGGCG

31981 + + + + + + 32040

GTTGCGGCGCCCGTGGTGACTCAAGCCCCCCCACAAGTACAAGCTCACGTGGCGGGCCGC
3 NAAGTTEFGGVFMFECTARR

CGAGGCCTGCCACATCATCGAACCCTCGTGCATCGAGGAGGTGCTCGACCCGGTGACGGA

32041 + + + + + + 32100

GCTCCGGACGGTGTAGTAGCTTGGGAGCACGTAGCTCCTCCACGAGCTGGGCCACTGCCT
3 EACHIIEPSCIEEVLDPVTE-

ACAGCCCGTCGGCTACGGCGAGGAGGGCGTCCGAGTCACCACCGGGCTGAACCGTGAGGG

32101 + + + + + + 32160

TGTCGGGCAGCCGATGCCGCTCCTCCCGCAGGCTCAGTGGTGGCCCGACTTGGCACTCCC
3 QPVGYGEEGVRVTTGLNREG-

GATGCAGCTCTTCCGGCACTGGACCGAGGACGTCGTGGTCAAGCGGCCCCACACCGAGTG

32161 + + + + + + 32220

CTACGTCGAGAAGGCCGTGACCTGGCTCCTGCAGCACCAGTTCGCCGGGGTGTGGCTCAC
3 MQL'FRHWTEDVVVKRPHTEC

CGGCTGCGGCCGGACGTGGGACTTCTACGACGGCGGCATCCTTCGGCGCGTGGACGACAT

32221 + + + + + + 32280

GCCGACGCCGGCCTGCACCCTGAAGATGCTGCCGCCGTAGGAAGCCGCGCACCTGCTGTA
3 GCGRTWDFYDGGILRRVDDM-

GCGCAAGATACGCGGGGTCTCGATCACCCCGGTGATGATCGAGGATGTGCTGCGCGGCTT

32281 + + + + + + 32340

CGCGTTCTATGCGCCCCAGAGCTAGTGGGGCCACTACTAGCTCCTACACGACGCGCCGAA
3 RKIRGVS ITPVMIEDVLRGF-

CGACGAGGTGAACGAGTTCCACTCGTCCATCCGGACCGTCCGCGGACTCGATACGATCCA

32341 + + + + + + 32400

GCTGCTCCACTTGCTCAAGGTGAGCAGGTAGGCCTGGCAGGCGCCTGAGCTATGCTAGGT
3 DEVNEFHSS IRTVRGLDTIH-

CGTCAAGGTCGAGGCGGGAGACATCTCGGGTGAGGCGGCCGAGAGCCTGTGCGGCCGCAT

32401 + + + + + + 32460

GCAGTTCCAGCTCCGCCCTCTGTAGAGCCCACTCCGGCGGCTCTCGGACACGCCGGCGTA
3 VKVEAGDISGEAAESLCGRI

CACCGAGGAGTTCAAGCGTGAGATAGGCATACGGCCCCAGGTGGAGCTGACCCCCGCGGG

32461 + + + + + + 32520

GTGGCTCCTCAAGTTCGCACTCTATCCGTATGCCGGGGTCCACCTCGACTGGGGGCGCCC
3 TEEFKRE IGIRPQVELTPAG-

CAGCCTCCCCCGATCGAAGTGGAAGGCGGCACGACTTCATGACGAGCGCGAACTCGCCCC

32521 + + + + + + 32580

GTCGGAGGGGGCTAGCTTCACCTTCCGCCGTGCTGAAGTACTGCTCGCGCTTGAGCGGGG
3 SLPRSKWKAARLHDERELAP

TCAGGCCTGAGCAGGTGGAGCAGCTCCTGGTGAGCTACCGGAGCCTGGGCCTGCTGGAGC

32581 + + + + + + 32640

AGTCCGGACTCGTCCACCTCGTCGAGGACCACTCGATGGCCTCGGACCCGGACGACCTCG
3-* Q A * -

AGAGCTGCGCGGTCCCGGCCGTGCTCGCCGCGGTCAGGGCCGCCCGTGCGGAACTCCGTA

32641 + + + + + + 32700

TCTCGACGCGCCAGGGCCGGCACGAGCGGCGCCAGTCCCGGCGGGCACGCCTTGAGGCAT

41

TCGCCCTGGACGGCCAGGGCGTGGAGTTCGAGTACTACCGGGGGCACGACGACAGCCTCG

32701 + + + + + + 32760

AGCGGGACCTGCCGGTCCCGCACCTCAAGCTCATGATGGCCCCCGTGCTGCTGTCGGAGC

TGGCCTGAACCCACCCCCGGTCCGCCGGGTCAGACGAAAGGGAGACCGGTGCCCCACGGT

32761 + + + + + + 32820

ACCGGACTTGGGTGGGGGCCAGGCGGCCCAGTCTGCTTTCCCTCTGGCCACGGGGTGCCA
> M P H G

GCAGAGCGCGAAGCGAGCCCGGCCGAGGAGAGCGCCGGCACCCGGCCGCTGACCGGCGAG

32821 + + + + + + 32880

CGTCTCGCGCTTCGCTCGGGCCGGCTCCTCTCGCGGCCGTGGGCCGGCGACTGGCCGCTC
AEREAS PAEESAGTRPLTGE

GAGTATCTGGAGAGCCTGCGGGACGCGCGGGAGGTGTACCTCGACGGCAGCCGCGTCAAG

32881 + + + + + + 32940

CTCATAGACCTCTCGGACGCCCTGCGCGCCCTCCACATGGAGCTGCCGTCGGCGCAGTTC
EYLESLRDAREVYLDGSRVK

GACGTCACCGCGCATCCCGCGTTCCACAACCCGGCCCGGATGACGGCCCGGCTGTACGAC

32941 + + + + + + 33000

CTGCAGTGGCGCGTAGGGCGCAAGGTGTTGGGCCGGGCCTACTGCCGGGCCGACATGCTG
DVTAHPAFHNPARMTARLYD

AGCCTGCACGACCCCGCCCAGAAAGCGGTCCTGACGGCGCCCACCGATGCCGGTGACGGT

33001 + + + + + + 33060

TCGGACGTGCTGGGGCGGGTCTTTCGCCAGGACTGCCGCGGGTGGCTACGGCCACTGCCA
SLHDPAQKAVLTAPTDAGDG

TTCACCCACCGCTTCTTCACCGCACCGCGCAGCGTCGACGACCTGGTCAAGGACCAGGCC

33061 + + + + + + 33120

AAGTGGGTGGCGAAGAAGTGGCGTGGCGCGTCGCAGCTGCTGGACCAGTTCCTGGTCCGG
FTHRFFTAPRSVDDLVKDQA

GCCATCGCATCCTGGGCGCGCAAGAGCTACGGCTGGATGGGGCGCAGCCCCGACTACAAG

33121 + + + + + + 33180

CGGTAGCGTAGGACCCGCGCGTTCTCGATGCCGACCTACCCCGCGTCGGGGCTGATGTTC
AIASWARKSYGWMGRS PDYK

GCGTCGTTCCTCGGCACGCTGGGGGCCAACGCCGACTTCTACGAGCCCTTCGCGGACAAC

33181 + + + + + + 33240

CGCAGCAAGGAGCCGTGCGACCCCCGGTTGCGGCTGAAGATGCTCGGGAAGCGCCTGTTG
AS FLGTLGANADFYE P FADN

GCCCGGCGCTGGTACCGGGAGTCGCAGGAGAAGGTGCTGTACTGGAACCATGCCTTCCTT

33241 + + + + + + 33300

CGGGCCGCGACCATGGCCCTCAGCGTCCTCTTCCACGACATGACCTTGGTACGGAAGGAA
ARRWYRESQEKVLYWNHAFL

CACCCGCCGGTCGACCGCTCGCTGCCCGCCGACGAGGTGGGCGACGTCTTCATCCACGTC

33301 + + + + + + 33360

GTGGGCGGCCAGCTGGCGAGCGACGGGCGGCTGCTCCACCCGCTGCAGAAGTAGGTGCAG
HPPVDRSLPADEVGDVFIHV

GAGCGGGAGACCGACGCGGGCCTGGTGGTGAGCGGGGCCAAGGTCGTCGCGACCGGATCG

33361 + + + + + + 33420

CTCGCCCTCTGGCTGCGCCCGGACCACCACTCGCCCCGGTTCCAGCAGCGCTGGCCTAGC
ERETDAGLVVSGAKVVATGS

GCCCTCACCCACGCGGCGTTCATCTCGCACTGGGGACTTCCCATCAAGGACCGGAAGTTC

33421 + + + + + + 33480

CGGGAGTGGGTGCGCCGCAAGTAGAGCGTGACCCCTGAAGGGTAGTTCCTGGCCTTCAAG
ALTHAAFI SHWGLP I KDRKF

GCCCTGGTGGCCACCGTGCCGATGGACGCGGACGGCCTCAAGGTGATCTGCCGTCCCTCC

42

33481 + + + + + + 33540

CGGGACCACCGGTGGCACGGCTACCTGCGCCTGCCGGAGTTCCACTAGACGGCAGGGAGG
ALVATVPMDADGLKVI CRPS

TACTCCGCAAACGCGGCGACCACGGGCAGCCCGTTCGACAACCCGCTGTCCTCACGGCTG

33541 + + + + + + 33600

ATGAGGCGTTTGCGCCGCTGGTGCCCGTCGGGCAAGCTGTTGGGCGACAGGAGTGCCGAC
YSANAATTGS PFDNPLSSRL

GACGAGAACGACGCCATCCTCGTACTCGACCAGGTGCTGATCCCCTGGGAGAACGTGTTC

33601 + + + + + + 33660

CTGCTCTTGCTGCGGTAGGAGCATGAGCTGGTCCACGACTAGGGGACCCTCTTGCACAAG
DENDAILVLDQVLI PWENVF

GTCTACGGCAACCTGGGCAAGGTACATCTCCTCGCCGGACAGTCCGGGATGATCGAACGC

33661 + + + + + + 33720

CAGATGCCGTTGGACCCGTTCCATGTAGAGGAGCGGCCTGTCAGGCCCTACTAGCTTGCG
VYGNLGKVHLLAGQSGMIER

GCCACCTTCCACGGGTGCACCCGGCTCGCCGTGAAGCTGGAGTTCATCGCCGGGCTGCTG

33721 + + + + + + 33780

CGGTGGAAGGTGCCCACGTGGGCCGAGCGGCACTTCGACCTCAAGTAGCGGCCCGACGAC
ATFHGCTRLAVKLEFIAGLL

GCCAAGGCGCTGGACATCACCGGGGCGAAGGACTTCCGCGGTGTGCAGACCCGGCTCGGA

33781 + + + + + + 33840

CGGTTCCGCGACCTGTAGTGGCCCCGCTTCCTGAAGGCGCCACACGTCTGGGCCGAGCCT
AKALDITGAKDFRGVQTRLG

GAAGTCCTGGCCTGGCGCAACCTCTTCTGGTCACTGTCGGACGCGGCGGCCCGCAACCCC

33841 + + + + + + 33900

CTTCAGGACCGGACCGCGTTGGAGAAGACCAGTGACAGCCTGCGCCGCCGGGCGTTGGGG
EVLAWRNLFWSLSDAAARNP

GTCCCCTGGAAGAACGGCACGCTCCTGCCCAACCCTCAGGCGGGTATGGCCTACCGCTGG

33901 + + + + + + 33960

CAGGGGACCTTCTTGCCGTGCGAGGACGGGTTGGGAGTCCGCCCATACCGGATGGCGACC
VPWKNGTLLPNPQAGMAYRW

TTCATGCAGATCGGCTACCCGCGGGTCCTGGAGATCGTCCAACAGGACGTGGCCAGCGGC

33961 + + + + + + 34020

AAGTACGTCTAGCCGATGGGCGCCCAGGACCTCTAGCAGGTTGTCCTGCACCGGTCGCCG
FMQIGYPRVLEIVQQDVASG

CTCATGTACGTCAACTCCTCCACGGAGGACTTCCGCAACCCCGAGACCGGCCCCTACTTG

34021 + + + + + + 34080

GAGTACATGCAGTTGAGGAGGTGCCTCCTGAAGGCGTTGGGGCTCTGGCCGGGGATGAAC
LMYVNSSTEDFRNPETGPYL

GAGAAGTACCTCCGGGGCAGCGACGGCGCAGGCGCCGTCGAGCGTGTCAAGGTGATGAAG

34081 + + + + + + 34140

CTCTTCATGGAGGCCCCGTCGCTGCCGCGTCCGCGGCAGCTCGCACAGTTCCACTACTTC
EKYLRGSDGAGAVERVKVMK

CTGCTGTGGGACGCGGTGGGATCCGACTTCGGCGGCCGGCACGAACTCTACGAGCGGAAC

34141 + + + + + + 34200

GACGACACCCTGCGCCACCCTAGGCTGAAGCCGCCGGCCGTGCTTGAGATGCTCGCCTTG
LLWDAVGSDFGGRHELYERN

TACTCCGGGAACCACGAGAACACCCGGATCGAGTTGCTGCTGTCGCAGACGGCGAGCGGC

34201 + + + + + + 34260

ATGAGGCCCTTGGTGCTCTTGTGGGCCTAGCTCAACGACGACAGCGTCTGCCGCTCGCCG
YSGNHENTRIELLLSQTASG

AAACTGGACTCGTACATGGACTTCGCCCAGGCATGCATGGACGAGTACGACCTGGACGGC
34261 + + + + + + 34320

43

TTTGACCTGAGCATGTACCTGAAGCGGGTCCGTACGTACCTGCTCATGCTGGACCTGCCG
4 KLDSYMDFAQACMDEYDLDG

TGGACCGCTCCCGACCTGGAGTCGTTTCACGCGATGCGTTCCGCCTCCCGCGACCTTCTC

34321 + + + + + + 34380

ACCTGGCGAGGGCTGGACCTCAGCAAAGTGCGCTACGCAAGGCGGAGGGCGCTGGAAGAG
4 WTAPDLESFHAMRSASRDLL

GGAGGGCTGTAGTTCCCCGACGGTGTACTGCGGCCCCCGATCCGGGGGCCGCAGTACACC

34381 + + + + + + 34440

CCTCCCGACATCAAGGGGCTGCCACATGACGCCGGGGGCTAGGCCCCCGGCGTCATGTGG
4-* G G L * -

GTCGGGGCGGCTGGTGCTCAGCCGCGCAGGAATCCGATGAGCTCGGGGGCGAGCTTCTTG

34441 + + + + + + 34500

CAGCCCCGCCGACCACGAGTCGGCGCGTCCTTAGGCTACTCGAGCCCCCGCTCGAAGAAC
22-* *GRLFGILEPALKK-

GGCGCCATGGCGACGGCACCGTGGTTGAGCCCGTTCAGGGTGCGGTGGCTCGCGTCGGGG

34501 + + + + + + 34560

CCGCGGTACCGCTGCCGTGGCACCAACTCGGGCAAGTCCCACGCCACCGAGCGCAGCCCC
22 PAMAVAGHNLGNLTRHSADP-

AGGACTCCGGTGAGTTCCTTCGCGGCACGCTGGAAACCGTCGGGGCTCTTGGAACCGGTC

34561 + + + + + + 34620

TCCTGAGGCCACTCAAGGAAGCGCCGTGCGACCTTTGGCAGCCCCGAGAACCTTGGCCAG
22 LVGTLEKAARQFGDPSKSGT-

AGCACCAGGGTCGGGGCCGACGCCGCCGACCACGGCTCGGCGGGGAGCGGCTTGCCCTGC

34621 + + + + + + 34680

TCGTGGTCCCAGCCCCGGCTGCGGCGGCTGGTGCCGAGCCGCCCCTCGCCGAACGGGACG
22 LVLTPASAASWPEAPLPKGQ

TGGGTGTCGCCCATCACCGCGATGTCGTAGGGAAGCGTGTTGGCCAGACCCTTGAGGTTG

34681 + + + + + + 34740

ACCCACAGCGGGTAGTGGCGCTACAGCATCCCTTCGCACAACCGGTCTGGGAACTCCAAC
22 QTDGMVAIDYPLTNALGKLN-

GACCAGACACCGGGCATCAGGCGCATGGCGCCGACCATGAAGGAGGGCATGCCCTGTGCC

34741 + + + + + -t- 34800

CTGGTCTGTGGCCCGTAGTCCGCGTACCGCGGCTGGTACTTCCTCCCGTACGGGACACGG
22 SWVGPMLRMAGVMFS PMGQA-

TTGACCATGAAGGCCTTGACCGCGTCGCTGCGTCGGTCCTCCGCCAGAAGGCTGTCGATC

34801 + + + + + + 34860

AACTGGTACTTCCGGAACTGGCGCAGCGACGCAGCCAGGAGGCGGTCTTCCGACAGCTAG
22 KVMFAKVADSRRDEALLSDI

TGACCGCCGAAGCCGGCGGGCGGGCCGAAGCCGTCCGAGGTGACGGAGAACGGCGGCTCG

34861 + + + + + + 34920

ACTGGCGGCTTCGGCCGCCCGCCCGGCTTCGGCAGGCTCCACTGCCTCTTGCCGCCGAGC
22 QGGFGAPPGFGDSTVS FPPE

TAGACCGCGAGCTTGTTCACCTTCAGGCCGGCGGCGGCGGCTCGCAGGGCGAGCACCGCG

34921 + + + + + + 34980

ATCTGGCGCTCGAACAAGTGGAAGTCCGGCCGCCGCCGCCGAGCGTCCCGCTCGTGGCGC
22 YVALKNVKLGAAAARLALVA

CCGGAAGAGCTGCCGAACAGGGAGGCCGAACCGCCGACCTGGTCGATCAGCGCCGCGATG

34981 + + + + + + 35040

GGCCTTCTCGACGGCTTGTCCCTCCGGCTTGGCGGCTGGACCAGCTAGTCGCGGCGCTAC
22 GSSSGFLSASGGVQDILAAI-

TCCTCGATCTCGCGCTCGACCGCGTACGCCGGACCGTCGGCGCTGGCGCCGCGGCCCCGA

35041 + + + + + + 35100

AGGAGCTAGAGCGCGAGCTGGCGCATGCGGCCTGGCAGCCGCGACCGCGGCGCCGGGGCT

44

22 DE I EREVAYAP, GDASAGRGR

CGGTCGTAGTTGACGACCGTGAAGTGCTCGGCGAGGAGACCGGCGAGCTTCTTGGCGTCG

35101 + + + + + + 35160

GCCAGCATCAACTGCTGGCACTTCACGAGCCGCTCCTCTGGCCGCTCGAAGAACCGCAGC
22 RDYNVVTFHEALLGALKKAD

GAGCGGTCGGCCAAGGCGGAGGCCACCAGGATCACCGCCGGCCCCTCGCCCGACTTGTCG

35161 + + + + + + 35220

CTCGCCAGCCGGTTCCGCCTCCGGTGGTCCTAGTGGCGGCCGGGGAGCGGGCTGAACAGC
22 SRDALASAVLIVAPGEGSKD-

AAGGCGATCGTGGTGCCGTCGGCCGATACCGTCGTTGATTCCACCTTGGCTGCTTTCTCA

35221 + + + + + + 35280

TTCCGCTAGCACCACGGCAGCCGGCTATGGCAGCAACTAAGGTGGAACCGACGAAAGAGT
22 F IATTGDASVTTSEVKAAKE

CGGGTTGAAGACATAGCTTCCCTCAGATCACATTGTGGGGCGTGCTGCCGACAGTGGAGA

35281 + + + + + + 35340

GCCCAACTTCTGTATCGAAGGGAGTCTAGTGTAACACCCCGCACGACGGCTGTCACCTCT
22-< R T S S M -

CCGGCGTCCGGAGGAAAAGTAATCGGTCCTGCCAGAATTGGGGGTTCCGGAGGGCACGCC

35341 + + + + + + 35400

GGCCGCAGGCCTCCTTTTCATTAGCCAGGACGGTCTTAACCCCCAAGGCCTCCCGTGCGG

GACCGCTGCACGACGGCGCGCCCCGACCTTCCGGACATTGTCGTGCCCTCAGATGTGTTT

35401 + + + + + + 35460

CTGGCGACGTGCTGCCGCGCGGGGCTGGAAGGCCTGTAACAGCACGGGAGTCTACACAAA

CGCATCTTCAGGAGTGCTCAGTGATCCGTGAGGTGAGAAAGGGACGGTGGTCCGGTCAGT

35461 + + + + + + 35520

GCGTAGAAGTCCTCACGAGTCACTAGGCACTCCACTCTTTCCCTGCCACCAGGCCAGTCA
18-* *

CGTTGCCGCGCGGGCTGTTCTGGTAAGCGGCCAGACGCCACTGCCCGTCCTGTTCGACGG

35521 + + + + + + 35580

GCAACGGCGCGCCCGACAAGACCATTCGCCGGTCTGCGGTGACGGGCAGGACAAGCTGCC
18 DNGRPSNQYAALRWQGDQEV

CCAGCCAGGAGGCCCGGACGGCGCCGTCGCCGCTCGCCTCGGTCTCCCCCGGGGCGAGGA

35581 + + + + + + 35640

GGTCGGTCCTCCGGGCCTGCCGCGGCAGCGGCGAGCGGAGCCAGAGGGGGCCCCGCTCCT
18 ALWSARVAGDGSAETEGPAL

TGCCGCCCTCGGTGATGAGCAGGGCGATGCCGTCGCCGAGCAGGCGCGCGTCGATGGGGC

35641 + + + + + + 35700

ACGGCGGGAGCCACTACTCGTCCCGCTACGGCAGCGGCTCGTCCGCGCGCAGCTACCCCG
18 IGGETILLAIGDGLLRADIP

TGCCGATGACACGGGTGCCCTTGTACGGGCCCGCGAAGGCGGCCGCCATGTGGGTGCGGA

35701 + + + + + + 35760

ACGGCTACTGTGCCCACGGGAACATGCCCGGGCGCTTCCGCCGGCGGTACACCCACGCCT
18 SGIVRTGKYPGAFAAAMHTR

TGTTCTCGCGGCCCTTGCGGAAGAGGCCGGGGAGGATCATCGTCCCGTCCTCGGCGAAGA

35761 + + + + + + 35820

ACAAGAGCGCCGGGAACGCCTTCTCCGGCCCCTCCTAGTAGCAGGGCAGGAGCCGCTTCT
18 INERGKRFLGPLIMTGDEAF

CGTCGGCGAACCGGTCGGCGTCGTGGTCGGCCCAGGCGGCCACGATGCGCGCCGGCAGAG

35821 + + + + + + 35880

GCAGCCGCTTGGCCAGCCGCAGCACCAGCCGGGTCCGCCGGTGCTACGCGCGGCCGTCTC
18 VDAFRDADHDAWAAVIRAPL

CGGCTACCGCTGCCAGGGCGGCGTCGGGAGCGGAGGTGGTCGAGTCGGTGCTGGTCATAT

45

35881 + + + + + + 35940

GCCGATGGCGACGGTCCCGCCGCAGCCCTCGCCTCCACCAGCTCAGCCACGACCAGTATA
-< AAVAALAADPASTTSDTSTM

CGCGGTTCCCGTCCGTTGGTTGGCGGTTTCGGCACGGCCCGCAGCCCTGCCCGAGCCCGA

3594I + + + + + + 36000

GCGCCAAGGGCAGGCAACCAACCGCCAAAGCCGTGCCGGGCGTCGGGACGGGCTCGGGCT

CGCTGGCAGGCGGCCCCGTCATCAGGCATCTCCTGCGTTGCGCCCCACGCCAGTCACTTC

36001 + + + + + + 36060

GCGACCGTCCGCCGGGGCAGTAGTCCGTAGAGGACGCAACGCGGGGTGCGGTCAGTGAAG

ACGGCCAGAACAAGTCGCGCATTCTGGAAGAAGCTGAGGCCCGCGACCCGGTGCGACGAT

36 061 + + + + + + 36120

TGCCGGTCTTGTTCAGCGCGTAAGACCTTCTTCGACTCCGGGCGCTGGGCCACGCTGCTA

CTGCGGTGTCACGGAGTTCGCACACGTTTACGCACGGAGGCTCGATGCCCGCTGTCAATG

36121 + + + + + + 36180

GACGCCACAGTGCCTCAAGCGTGTGCAAATGCGTGCCTCCGAGCTACGGGCGACAGTTAC
> MPAVNG-

GATCGGTGCAGTCAGGCCAGTCGCACCGACGCTCCGTCGTGGCGACGGTGGTGGGCAACT

36181 + + + + + + 36240

CTAGCCACGTCAGTCCGGTCAGCGTGGCTGCGAGGCAGCACCGCTGCCACCACCCGTTGA
SVQSGQ SHRRSVVATVVGNF-

TCGTGGAGTCGTTCGACTGGCTCGCCTACGGGCTCTTCGCTCCTCTCTTCGCGGCTCAGT

36241 + + + + + + 36300

AGCACCTCAGCAAGCTGACCGAGCGGATGCCCGAGAAGCGAGGAGAGAAGCGCCGAGTCA
VESFDWLAYGLFAPLFAAQF-

TCTTCCCCTCGTCCAACCAGTTCACCTCCCTGCTCGGCGCGTTCGCGGTCTTCGGCACGG

36301 + + + + + + 36360

AGAAGGGGAGCAGGTTGGTCAAGTGGAGGGACGAGCCGCGCAAGCGCCAGAAGCCGTGCC
FPSSNQFTSLLGAFAVFGTG-

GCATGCTCTTCCGGCCGATCGGCGGGGTCCTGCTGGGCCGCCTCGCCGACCGGCGCGGCC

36361 + + + + + + 36420

CGTACGAGAAGGCCGGCTAGCCGCCCCAGGACGACCCGGCGGAGCGGCTGGCCGCGCCGG
ML FR P I GGVLLGRLADRRGR-

GGCGCCCCGCCCTGATGCTGGCGATCGGACTGATGACCGGCGGCTCGACCCTGATCGCCG

36421 + + + + + + 36480

CCGCGGGGCGGGACTACGACCGCTAGCCTGACTACTGGCCGCCGAGCTGGGACTAGCGGC
RPALMLAI GLMTGGSTL I AV-

TCGTCCCCACCTACGAGCACATCGGGATCCTCGCCCCGCTGCTTCTGCTGCTCGCCCGGC

36481 + + + + + + 36540

AGCAGGGGTGGATGCTCGTGTAGCCCTAGGAGCGGGGCGACGAAGACGACGAGCGGGCCG
VPTYEHIGILAPLLLLLARL-

TCGCCCAGGGAGTCTCCTCGGGCGGGGAATGGACAGCGGCGGCCACCTACCTGATGGAGA

36541 + + + + + + 36600

AGCGGGTCCCTCAGAGGAGCCCGCCCCTTACCTGTCGCCGCCGGTGGATGGACTACCTCT
AQGVS SGGEWTAAATYLME I -

TCGCGCCGAAGAACCGCCGGTGCCTCTACAGCAGCCTCTTCTCCGTGACGACCATGGCGG

36601 + + + + + + 36660

AGCGCGGCTTCTTGGCGGCCACGGAGATGTCGTCGGAGAAGAGGCACTGCTGGTACCGCC
APKNRRCLYSSLFSVTTMAG-

GCCCCTTCGTCGCATCGCTGCTGGGCGCGGGCCTCGGCGTGTGGCTGGGAACCGCGACGA

36661 + + + + + + 36720

CGGGGAAGCAGCGTAGCGACGACCCGCGCCCGGAGCCGCACACCGACCCTTGGCGCTGCT
PFVASLLGAGLGVWLGTATM-

46

TGGAGGCCTGGGGCTGGCGGGTGCCGTTCCTCCTCGGCGGCGTCTTCGGCGTGATCCTGC

36721 + + + + + + 36780

ACCTCCGGACCCCGACCGCCCACGGCAAGGAGGAGCCGCCGCAGAAGCCGCACTAGGACG
5 EAWGWRVPFLLGGVFGVILL-

TGTTCCTGCGCCGTCGGCTCACCGAGACCGAGGTCTTCCGCCGGGAGGTGCGGCCCCGGG

36781 + + + + + + 36840

ACAAGGACGCGGCAGCCGAGTGGCTCTGGCTCCAGAAGGCGGCCCTCCACGCCGGGGCCC
5 FLRRRLTETEVFRREVRPRA-

CCCGGCGCGGCTCACTGGGCCAGCTGATCGGAGCCCACCGCCCCCAGGTGCTGCTGGCCG

36841 + + + + + + 36900

GGGCCGCGCCGAGTGACCCGGTCGACTAGCCTCGGGTGGCGGGGGTCCACGACGACCGGC
5 RRGSLGQL IGAHRPQVLLAV-

TGATGCTGGTGGCCGGACTGGGCGTCATCGGCGGAACGTGGTCGACCGCGGTCCCGGCGA

36901 + + + + + + 36960

ACTACGACCACCGGCCTGACCCGCAGTAGCCGCCTTGCACCAGCTGGCGCCAGGGCCGCT
5 MLVAGLGVI GGTW S TAVPAM-

TGGGCCACCGTCTGATCGGCTCGCAGACGATGTTCTGGGTGGTGGTCTGTGTGACCGGCT

36961 + + + + + + 37020

ACC CGGTGGCAGACTAGCCGAGCGTCTGCTACAAGACCCACCACCAGACACACTGGC CGA
5 GHRL I GSQTMFWVVVCVTGS-

CGGTCATCCTGCTGCAGGTACCCATAGGGCTGCTCGCCGACCGGGTGGAACCGGGCAGGT

37021 + + + + + + 37080

GCCAGTAGGACGACGTCCATGGGTATCCCGACGAGCGGCTGGCCCACCTTGGCCCGTCCA
5 VI LLQVP IGLLADRVE PGRF-

TCCTGATCGTCTCCAGCGTCGTCTTCGCCGCTGTGGGCTCGTACGCCTACCTCACCGTCC

37081 + + + + + + 37140

AGGACTAGCAGAGGTCGCAGCAGAAGCGGCGACACCCGAGCATGCGGATGGAGTGGCAGG
5 LIVSSVVFAAVGSYAYLTVQ-

AGGACTCCTTCGCGAGCCTGGCGTTCACGTACAGCACCGGAGTGATCTTCCTCGGCTGCG

37141 + + + + + + 37200

TCCTGAGGAAGCGCTCGGACCGCAAGTGCATGTCGTGGCCTCACTAGAAGGAGCCGACGC
5 DS FASLAFTYSTGVI FLGCV-

TCACCATGGTGCTGCCGAAGATGCTCTCCAGAATCTTCCCTCCGCAGATACGCGGCCTGG

37201 + + + + + + 37260

AGTGGTACCACGACGGCTTCTACGAGAGGTCTTAGAAGGGAGGCGTCTATGCGCCGGACC
5 TMVLPKMLSRI FPPQI RGLG-

GCATCGGGCTGCCGCACGCCTCGACCACCGCACTCCTCGGCGGGGCGGGGCCACTGCTGG

37261 + + + + + + 37320

CGTAGCCCGACGGCGTGCGGAGCTGGTGGCGTGAGGAGCCGCCCCGCCCCGGTGACGACC
5 I GLPHAS TTALLGGAG PLLA-

CCGCCTACTCCGACGAGCGAGGCGCCTCGGGCTGGTTCATCGCCGCCGTGATGGCCGCGG

37321 + + + + + + 37380

GGCGGATGAGGCTGCTCGCTCCGCGGAGCCCGACCAAGTAGCGGCGGCACTACCGGCGCC
5 AYS DERGASGWF I AAVMAAV-

TCCTGCTCGCCTGGCCGGCCACCCTGTGGGAGCGACGGCTGTTCCGCGCCCGGACGGCCC

37381 + -f + + + + 37440

AGGACGAGCGGACCGGCCGGTGGGACACCCTCGCTGCCGACAAGGCGCGGGCCTGCCGGG
5 LLAWPATLWERRLFRARTAP-

CGGGAAGCGAGCCGGTTCCCGAATCCGCCGTCGCCCGCCCCGTCGGGTGACCGTCCGCAC

37441 + + + + + + 37500

GCCCTTCGCTCGGCCAAGGGCTTAGGCGGCAGCGGGCGGGGCAGCCCACTGGCAGGCGTG
5-* GSEPVPESAVARPVG*-

47

TTCTGCATCCCGTCCGGCACCGAGCGCCGGCGACCTTCCCGACTGAGAGGTTGACATCAT

37501 + + + + + + 37560

AAGACGTAGGGCAGGCCGTGGCTCGCGGCCGCTGGAAGGGCTGACTCTCCAACTGTAGTA
23-> M -

GACGACGTCCGACACCACCGACCGGTCCCAGGACGGCGTGCCGCCGCTCTCCTTCCACCA

37561 + + + + + + 37620

CTGCTGCAGGCTGTGGTGGCTGGCCAGGGTCCTGCCGCACGGCGGCGAGAGGAAGGTGGT
23 TTSDTTDRSQDGVPPLSFHQ-

GGAGTTCCTGTGCATGTTCGACAGCGGGAACGACGGCGCCGACGTGGGGCCGTTCGGCCC

37621 + + + + + + 37680

CCTCAAGGACACGTACAAGCTGTCGCCCTTGCTGCCGCGGCTGCACCCCGGCAAGCCGGG
23 EFLCMFDSGNDGADVGPFGP-

CATGTACCACATCGTCGGAGCCTGGCGGCTGACCGGCGGGATCGACGAGGAGACCCTGCG

37681 + + + + + + 37740

GTACATGGTGTAGCAGCCTCGGACCGCCGACTGGCCGCCCTAGCTGCTCCTCTGGGACGC
23 MYHIVGAWRLTGGIDEETLR

CGAGGCGCTGGGTGACGTCGTCGTGCGCCACGAGGCCCTGCGCACATCGCTGGTCCGCGA

37741 + + + + + + 37800

GCTCCGCGACCCACTGCAGCAGCACGCGGTGCTCCGGGACGCGTGTAGCGACCAGGCGCT
23 EALGDVVVRHEALRTSLVRE-

AGGTGGCACGCACCGGCCGGAGATCCTGCCTGCGGGGCCCGCCGCGCTGGAGGTCCGTGA

37801 + + + + + + 37860

TCCACCGTGCGTGGCCGGCCTCTAGGACGGACGCCCCGGGCGGCGCGACCTCCAGGCACT
23 GGTHRPEI LPAGPAALEVRD

TCTCGGCGACGTCGACGAGTCGGAGCGGGTGCGGCGCGGTGAGGAACTGCTCAACGAGGT

37861 + + + + + + 37920

AGAGCCGCTGCAGCTGCTCAGCCTCGCCCACGCCGCGCCACTCCTTGACGAGTTGCTCCA
23 LGDVDESERVRRGEELLNEV-

GGAGTCGACCGGTCTGAGCGTGCGGGAGCTGCCCCTGCTGCGGGCCGTGCTCGGACGCTT

37921 + + + + + + 37980

CCTCAGCTGGCCAGACTCGCACGCCCTCGACGGGGACGACGCCCGGCACGAGCCTGCGAA
23 ESTGLSVRELPLLRAVLGRF

CGACCAGAAGGACGCGGTGCTGGTCCTCATCGCCCACCACACCGCCGCGGACGCCTGGGC

37981 + + + + + + 38040

GCTGGTCTTCCTGCGCCACGACCAGGAGTAGCGGGTGGTGTGGCGGCGCCTGCGGACCCG
23 DQKDAVLVL IAHHTAADAWA

CATGCACGTCATCGCCCGCGACCTGCTCAACCTGTACGCCGCCAGGCGCGGGAACCCGGT

38041 + + + + + + 38100

GTACGTGCAGTAGCGGGCGCTGGACGAGTTGGACATGCGGCGGTCCGCGCCCTTGGGCCA
23 MHVIARDLLNLYAARRGNPV-

TCCCCCGCTCCCCGAGCCGGCCCAGCATGCCGAGTTCGCCCGCTGGGAGCGCGAGGCGGC

38101 + + + + + + 38160

AGGGGGCGAGGGGCTCGGCCGGGTCGTACGGCTCAAGCGGGCGACCCTCGCGCTCCGCCG
23 PPLPEPAQHAEFARWEREAA-

CGAGGCACCGCGGGTCGCGGTCTCGAAGGAATTCTGGCGCAAGCGCCTCCAGGGCGCGCG

38161 + + + + + + 38220

GCTCCGTGGCGCCCAGCGCCAGAGCTTCCTTAAGACCGCGTTCGCGGAGGTCCCGCGCGC
23 EAPRVAVS KEFWRKRLQGAR

GATCATCGGGCTGGAGACGGACATACCGCGCTCGGCGGGGCTGCCCAAGGGCACCGCGTG

38221 + + + + + + 38280

CTAGTAGCCCGACCTCTGCCTGTATGGCGCGAGCCGCCCCGACGGGTTCCCGTGGCGCAC
23 I IGLETD I PRSAGLPKGTAW

GCAGCGCTTCGCCGTACGCGGGGAACTGGCCGACGCCGTGGTGGAGTTCTCACGGGCCGC

48

38281 + + + + + + 38340

CGTCGCGAAGCGGCATGCGCCCCTTGACCGGCTGCGGCACCACCTCAAGAGTGCCCGGCG
23 QRFAVRGELADAVVEFSRAA-

CAAGTGCTCCCCGTTCATGACCATGTTCGCCGCCTACCAGGTGCTGCTGCACCGCAGGAC

38341 + + + + + + 38400

GTTCACGAGGGGCAAGTACTGGTACAAGCGGCGGATGGTCCACGACGACGTGGCGTCCTG
23 KCS PFMTMFAAYQVLLHRRT

GGGCGAGCTGGACATCACCGTGCCGACCTTCTCCGGGGGGCGCAACAACTCGCGGTTCGA

38401 + + + + + + 38460

CCCGCTCGACCTGTAGTGGCACGGCTGGAAGAGGCCCCCCGCGTTGTTGAGCGCCAAGCT
23 GELDITVPTFSGGRNNSRFE-

GGACACCGTCGGTTCCTTCATCAACTTCCTGCCGCTGCGTACCGACCTCTCCGGATGCGC

38461 + + + + + + 38520

C CTGTGGCAGCCAAGGAAGTAGTTGAAGGACGGCGACGCATGGCTGGAGAGGC CTACGCG
23 DTVGSFINFLPLRTDLSGCA-

ATCCTTCCGCGAGGTCGTGCTGCGCACCCGCACCACCTGCGGAGAGGCGTTCACCCACGA

38521 + + + + + + 38580

TAGGAAGGCGCTCCAGCACGACGCGTGGGCGTGGTGGACGCCTCTCCGCAAGTGGGTGCT
23 S FREVVLRTRTTCGEAFTHE

GCTGCCCTTCTCCCGGCTGATCCCGGAGGTGCCGGAGCTGATGGCGTCGGCGGCCTCCGA

38581 + + + + + + 38640

CGACGGGAAGAGGGCCGACTAGGGCCTCCACGGCCTCGACTACCGCAGCCGCCGGAGGCT
23 LPFSRLI PEVPELMASAASD

CAACCACCAGATCTCCGTCTTCCAGGCCGTGCACGCGCCCGCGTCCGAGGGGCCCGAGCA

38641 + + + + + + 38700

GTTGGTGGTCTAGAGGCAGAAGGTCCGGCACGTGCGCGGGCGCAGGCTCCCCGGGCTCGT
23 NHQI SVFQAVHAPASEGPEQ-

GGCCGGGGACCTGACGTACTCGAAGATCTGGGAGCGGCAGCTGTCGCAGGCGGAGGGCTC

38701 + + + + + + 38760

CCGGCCCCTGGACTGCATGAGCTTCTAGACCCTCGCCGTCGACAGCGTCCGCCTCCCGAG
23 AGDLTYSKIWERQLSQAEGS-

CGACATCCCCGACGGGGTGCTGTGGTCGATCCACATCGACCCCTCGGGCTCCATGGCCGG

38761 + + + + + + 38820

GCTGTAGGGGCTGCCCCACGACACCAGCTAGGTGTAGCTGGGGAGCCCGAGGTACCGGCC
23 DIPDGVLWSIHIDPSGSMAG-

CAGCCTCGGGTACAACACCAACCGCTTCAAGGACGAGACGATGGCGGCCTTCCTGGCCGA

38821 + + + + + + 38880

GTCGGAGCCCATGTTGTGGTTGGCGAAGTTCCTGCTCTGCTACCGCCGGAAGGACCGGCT
23 SLGYNTNRFKDETMAAFLAD-

CTACCTCGACGTGCTCGAGAACGCGGTGGCCCGGCCGGACGCCCCCTTCACCTCCTGAGA

38881 + + + + + + 38940

GATGGAGCTGCACGAGCTCTTGCGCCACCGGGCCGGCCTGCGGGGGAAGTGGAGGACTCT
23-* YLDVLENAVARPDAPFTS *-

CAGTTCCGGCGGCGGCGAACCCGCCCGAAGAAAGGAAAGCCAGTGTCCACCGTTTCCGAC

38941 + + + + + + 39000

GTCAAGGCCGCCGCCGCTTGGGCGGGCTTCTTTCCTTTCGGTCACAGGTGGCAAAGGCTG
26-> M S T V S D

ACAGCGGCCGGCTCCTCCCTGGAGGAGAAGGTCACCCGGATCTGGACGGGTGTTCTCGGC

39001 + + + + + + 39060

TGTCGCCGGCCGAGGAGGGACCTCCTCTTCCAGTGGGCCTAGACCTGCCCACAAGAGCCG
26 TAAGS SLEEKVTRIWTGVLG

ACGTCCGGTGAGGAAGGCGCGACGTTCATCGAGCTCGGAGGGCAGTCGGTCTCGGCCGTG
39061 + + + + + + 39120

49

TGCAGGCCACTCCTTCCGCGCTGCAAGTAGCTCGAGCCTCCCGTCAGCCAGAGCCGGCAC
TSGEEGATFIELGGQSVSAV

CGCATCGCCACGCGTATCCAGGAGGAGCTCGACATCTGGGTCGACATCGGCGTCCTCTTC

39121 + + + + + + 39180

GCGTAGCGGTGCGCATAGGTCCTCCTCGAGCTGTAGACCCAGCTGTAGCCGCAGGAGAAG
RIATRIQEELDIWVDIGVLF

GACGACCCGGATCTGCCTACCTTCATCGCGGCGGTCGTCCGGACGGCCGACGCCGCGGGC

39X81 + + + + + + 39240

CTGCTGGGCCTAGACGGATGGAAGTAGCGCCGCCAGCAGGCCTGCCGGCTGCGGCGCCCG
DDPDLPTF IAAVVRTADAAG

GGCGAGGGCTCCGGAACGCAGTGAGACTCGCCGGGCGCCGTCTCCCCGCGGCGCCCGGTT

39241 + + + + + + 39300

CCGCTCCCGAGGCCTTGCGTCACTCTGAGCGGCCCGCGGCAGAGGGGCGCCGCGGGCCAA

* GEGSGTQ*-

TCACATGGCTGAGGCGGTTCACCCGGTACCGGGTGAACCGCCTCAGCCATGTGAAACCGG

39301 + + + + + + 39360

AGTGTACCGACTCCGCCAAGTGGGCCATGGCCCACTTGGCGGAGTCGGTACACTTTGGCC

GCCTGGTCAGCGCAGCTGGATGTCCGTCTCCCGGGCGATCGCCCGGAGGAACTCGCCGCG

39361 + + + + + + 39420

CGGACCAGTCGCGTCGACCTACAGGCAGAGGGCCCGCTAGCGGGCCTCCTTGAGCGGCGC
.* *RLQIDTERAIARLFEGR-

GGACAGCGCGTCGGCGACCAGCTCGATGTCGTCGGCCATGTACCGGTCGACGCCCAGCGT

39421 + + + + + + 39480

CCTGTCGCGCAGCCGCTGGTCGAGCTACAGCAGCCGGTACATGGCCAGCTGCGGGTCGCA
S LADAVLE I DDAMYRDVGLT-

CGGAACCAGCCGGCGCACCGCTTCGTACGTGGCCTTCGCCGCCGGGCTCAAGCCGTCGAA

39481 + + + + + + 39540

GCCTTGGTCGGCCGCGTGGCGAAGCATGCACCGGAAGCGGCGGCCCGAGTTCGGCAGCTT
PVLRRVAEYTAKAAP S LGD F -

CCGGCCGGAGATGTCGACCGCCTGGGCGGCGGCCAGGTACTCCACCGCGAGGATCTTGTT

3954! + + + + + + 39600

GGCCGGCCTCTACAGCTGGCGGACCCGCCGCCGGTCCATGAGGTGGCGCTCCTAGAACAA
RG S I DVAQAAAL Y EVAL I KN-

GTTGTTCGACAGGACCCGGCGGGCGTTGCGGGCCGAGATCAGGCCCATGCTCACCACGTC

39601 + + + + + + 39660

CAACAAGCTGTCCTGGGCCGCCCGCAACGCCCGGCTCTAGTCCGGGTACGAGTGGTGCAG
NNSLVRRANRAS ILGMSVVD-

CTGGTTGTCGCCGTTGGACGGGACGCTCTGGGTGCTGGCCGGGCCGATCGTCCGGTTCTC

39661 + + + + + + 39720

GACCAACAGCGGCAACCTGCCCTGCGAGACCCACGACCGGCCCGGCTAGCAGGCCAAGAG

QNDGNS PVSQTSAPGITRNE-

GGCCACCAGTGCGGTGGCCGGGTACTGGGCGCCGGCGAATCCGCTGTGCAGCCCCGGGTC

39721 + + + + + + 39780

CCGGTGGTCACGCCACCGGCCCATGACCCGCGGCCGCTTAGGCGACACGTCGGGGCCCAG
AVLATAPYQAGAFGSHLGPD-

CCCGGAGACGAGGAACTCCGGGAGGCCGTAGCTGAGGTGCCGGTTCAGGACCCGGTTGAT

39781 + + + + + + 39840

GGGCGTCTGCTCCTTGAGGCCCTCCGGCATCGACTCCACGGCCAAGTCCTGGGCCAACTA
G SVLFE PLGYS LHRNLVRN I -

CTGCCGCTCGGCCAGGACGCCGAGCTGGGTGAGCGCGATGGTCACGAAGTCCATCGCGAA

39841 + + + + + + 39900

GACGGCGAGCCGGTCCTGCGGCTCGACCCACTCGCGCTACCAGTGCTTCAGGTAGCGCTT

50

24 QREALVGLQTLAI TVFDMAF-

CGCGATCGGCTGACCGTGGAAGTTCGCCCCGTGGAAGATCTCCTTGCCCTCGAAGAAGAG

399OI + + + + + + 39960

GCGCTAGCCGACTGGCACCTTCAAGCGGGGCACCTTCTAGAGGAACGGGAGCTTCTTCTC
24 AI PQGHFNAGHFI EKGEFFL-

CGGGTTGTCGTTGGCCGAGTTGAGCTCGATGCGCAGCTTGTGCCGCGCGTGGTACAAGGT

39961 + + + + + + 40020

GCCCAACAGCAACCGGCTCAACTCGAGCTACGCGTCGAACACGGCGCGCACCATGTTCCA
24 PNDNASNLE I RLKHRAHYLT-

GTCGCGCACCGCCCCGACGACCTGGGGGATGGCCCGCAGCGAGTAGGCCTTCTGCAGGTA

40021 + + + + + + 40080

CAGCGCGTGGCGGGGCTGCTGGACCCCCTACCGGGCGTCGCTCATCCGGAAGACGTCCAT
24 DRVAGVVQP IARL SYAKQLY-

GATCTCCGAGCGCTGGACGTCCTTGCCGGCCTCCTTGTCCTTCTGGAGTTCTCGGCGCAG

40081 + + + + + + 40140

CTAGAGGCTCGCGACCTGCAGGAACGGCCGGAGGAACAGGAAGACCTCAAGAGCCGCGTC
24 IESRQVDKGAEKDKQLERRL-

GTCGGCGTGCTCGACCGTCAGTCCGCTGCCCCGCATCAGGGCCCGCATGTTGGCGGCGGT

40141 + + + + + + 40200

CAGCCGCACGAGCTGGCAGTCAGGCGACGGGGCGTAGTCCCGGGCGTACAACCGCCGCCA
24 DAHEVTLGSGRMLARMNAAT-

GTCGATCTGGCCCTCGTGCGGGCGGGCTATGTCGTGCCCCTCCGCGAGGAAGGGGCTGGT

40201 + + + + + + 40260

CAGCTAGACCGGGAGCACGCCCGCCCGATACAGCACGGGGAGGCGCTCCTTCCCCGACCA
24 DIQGEHPRAIDHGEALFPST-

CGATCCGCGTACCGCCTCGATGAGCAGAGCCGTCACGATCTCGGCCTGCTGGGCCTGCTC

40261 + + + + + + 40320

GCTAGGCGCATGGCGGAGCTACTCGTCTCGGCAGTGCTAGAGCCGGACGACCCGGACGAG
24 SGRVAE I LLATV I EAQQAQE-

CAGGGCCCGTCCGACGACCAGGGAGCCCAGACCGGTCATCCCGGACGTGCCGTTGATCAG

40321 + + + + + + 40380

GTCCCGGGCAGGCTGCTGGTCCCTCGGGTCTGGCCAGTAGGGCCTGCACGGCAACTAGTC
24 LARGVVLSGLGTMGSTGNIL-

TGCGAGGCCCTCCTTGAAGCGCAGTTCGAGCGGCTCGATGCCCCGCTCGGCCAGCACCTG

40381 + + + + + + 40440

ACGCTCCGGGAGGAACTTCGCGTCAAGCTCGCCGAGCTACGGGGCGAGCCGGTCGTGGAC
24 ALGEKFRLELPE I GREALVQ-

GGCGGTCTCCACCGGCCGTCCGTCGCGCAGGACGTAGCCCTCTCCGATGAGGGTGCTCGC

40441 + + + + + + 40500

CCGCCAGAGGTGGCCGGCAGGCAGCGCGTCCTGCATCGGGAGAGGCTACTCCCACGAGCG
24 ATEVPRGDRLVYGEGI LTSA-

GACGTGGGAGAGGGGAGCCAGGTCGCCGCTCGCCCCGAGTGACCCGATCTCGGGTATGGC

40501 + + + + + + 40560

CTGCACCCTCTCCCCTCGGTCCAGCGGCGAGCGGGGCTCACTGGGCTAGAGCCCATACCG
24 VHSL PALDGSAGLSG I EP I A -

CGGGGTGATGCCCTCGTTCAGGTACTGCGCGAGGCGTTCGAGGATGATGGGGCGCACCGC

40561 + + + + + + 40620

GCCCCACTACGGGAGCAAGTCCATGACGCGCTCCGCAAGCTCCTACTACCCCGCGTGGCG

24 PT I GENLYQALREL I I PRVA-

GGAGTGGCCCTTGGCGAGGGTGTTCAGCCGGGCGGCGACGATCGCCCGCGCCTCGTCCTC

4062 i + + + + + + 40680

CCTCACCGGGAACCGCTCCCACAAGTCGGCCCGCCGCTGCTAGCGGGCGCGGAGCAGGAG

51

24 S HGKALTNLRAAV IARAEDE-

GGCGAACAGCGGACCGACTCCCGCGCTGTGGCTACGGACGAGATTGGTCTGCAGTTCGAC

40681 + + + + + + 40740

CCGCTTGTCGCCTGGCTGAGGGCGCGACACCGATGCCTGCTCTAACCAGACGTCAAGCTG
24 AFLPGVGAS HSRVLNTQLEV-

TTCCTTCGACTTGTCGACCTGCATGTAGATCATCTCGCCGTACCCGGTGGTCACCCCGTA

40741 + + + + + + 40800

AAGGAAGCTGAACAGCTGGACGTACATCTAGTAGAGCGGCATGGGCCACCAGTGGGGCAT
24 E KS KDVQMY I MEGYGTTVGY-

GATGGGGATGTTCTGTTCGGCGATCCCTTCGAAGATCTCCCGGCTCTTCTGGGCCTTCGC

40801 + + + + + + 40860

CTACCC CTACAAGACAAGC CGCTAGGGAAGCTTCTAGAGGGCCGAGAAGAC CCGGAAGCG
24 I PINQEAIGEFIERSKQAKA-

GATGGATTCGGCCGGTAGGTCGACCGTCGCGCGTTCCTCCGCGACGCGGCGTACGGCTTC

40861 + + + + + + 40920

CTACCTAAGCCGGCCATGCAGCTGGCAGCGCGCAAGGAGGCGCTGCGCCGCATGCCGAAG
24 I S EAPVDVTARE EAVRRVAE-

GACGGTCAGGGTCTCGCCGTCGACGGAAACCGGGACGATCTCGGTCTCGACTTGAGTCAA

40921 + + + + + + 40980

CTGCCAGTCCCAGAGCGGCAGCTGCCTTTGGCCCTGCTAGAGCCAGAGCTGAACTCAGTT

24 VTLTEGDVSVPVI ETEVQTL-

TGCCATCACTCCATGGGTAGCGGCCGAGGCCGGTGTACGACAGGTCAGGGGGTGGGTTCG

40981 + + + + + + 41040

ACGGTAGTGAGGTACCCATCGCCGGCTCCGGCCACATGCTGTCCAGTCCCCCACCCAAGC

24- < A M -

TGAGGCGCGGCTCAGCGGGTGAGCCGGGAGCGGTCCACCTTCCCCGCGGCGTTGCGCGGC

41041 + + + + + + 41100

ACTCCGCGCCGAGTCGCCCACTCGGCCCTCGCCAGGTGGAAGGGGCGCCGCAACGCGCCG

25- * *RTLRSRDVKGAANRP -

AGGCGTGAAGTCAGGCGGGTGAAGACGGCGGGCAGTGCGAGGGGGCCGAACTGGCCGCGC

41101 + + + + + + 41160

TCCGCACTTCAGTCCGCCCACTTCTGCCGCCCGTCACGCTCCCCCGGCTTGACCGGCGCG

25 LRSTLRTFVAPLALPGFQGR-

AGATGGGAACGCCAGGCCCGGATGTCCGCGCGCACGTCCTCCCGGCCCTCTCCTTGTGGC

41161 + + + + + + 41220

TCTACCCTTGCGGTCCGGGCCTACAGGCGCGCGTGCAGGAGGGCCGGGAGAGGAACACCG
25 LHSRWARIDARVDERGEGQP

ACCACGTACACGGCGAGGCGGGTCACCAGGCCCTGGCCGTTGACGTGGGGGAGGACCGCG

41221 + + + + + + 41280

TGGTGCATGTGCCGCTCCGCCCAGTGGTCCGGGACCGGCAACTGCACCCCCTCCTGGCGC
25 VVYVALRTVLGQGNVHPLVA-

CACTCCAGGACCGAGGGGTCACGGTTCAGCGCGGCCTCGATCTCGGTGAGTTCCAAGCGG

41281 + + + + + + 41340

GTGAGGTCCTGGCTCCCCAGTGCCAAGTCGCGCCGGAGCTAGAGCCACTCAAGGTTCGCC
25 CELVS PDRNLAAE I ETLELR-

TTCCCGAACAGCTTGACCTGGAAGTCCTTGCGGCCCCGGAATTCCAGGGCTCCGTCGAAC

41341 + + + + + + 41400

AAGGGCTTGTCGAACTGGACCTTCAGGAACGCCGGGGCCTTAAGGTCCCGAGGCAGCTTG
25 NGFLKVQFDKRGRFELAGDF-

CGTACCCGCGCCAGATCCCCGGTCCGGTACCACCGGTCACCGTCCGGGGCGAGGCCGGCG

41401 + + + + + + 41460

GCATGGGCGCGGTCTAGGGGCCAGGCCATGGTGGCCAGTGGCAGGCCCCGCTCCGGCCGC
25 RVRALDGTRYWRDGD PALGA-

52

AGGGGCGCGAACAGCGCGCTGTGGTCCGGGCCGCCCTCGACGGCGAGATAACCCGGCGTC

41461 + + + + + + 41520

TCCCCGCGCTTGTCGCGCGACACCAGGCCCGGCGGGAGCTGCCGCTCTATTGGGCCGCAG
LPAFLASHDPGGEVALYGPT-

ACGTACGGGGAGCGGATCACCAGTTCGCCGGTGACGCCGGCGGGGCTCGGCCGGTCGTCC

41521 + + + + + + 41580

TGCATGCCCCTCGCCTAGTGGTCAAGCGGCCACTGCGGCCGCCCCGAGCCGGCCAGCAGG
VYPSRIVLEGTVGAPSPRDD

GCGTCCACGACGAGTACCTGGCGGCCGGGGAGCGGGTACCCGATCGGGGCCGGGCCCGTG

41581 + + + + + + 41640

CGCAGGTGCTGCTCATGGACCGCCGGCCCCTCGCCCATGGGCTAGCCCCGGCCCGGGCAC
ADVVLVQRGPLPYGI PAPGT

ACCGGCCCGGTGATCTCGTGCCAGGTCGCGGCGATCGTCTCGGTGGGCCCGTAGAGGTTG

41641 + + + + + + 41700

TGGCCGGGCCACTAGAGCACGGTCCAGCGCCGCTAGCAGAGCCACCCGGGCATCTCCAAC
VPGTIEHWTAAITETPGYLN-

ATCAGGCGGGTCCGGGGCAGGGCCGCGCGCAGTCCGTCCACGAGTTCGCCGGGCAGCGCC

41701 + + + + + + 41760

TAGTCCGCCCAGGCCCCGTCCCGGCGCGCGTCAGGCAGGTGCTCAAGCGGCCCGTCGCGG
ILRTRPLAARLGDVLEGPLA-

TCGCCCATCAGGAGCAGGTGGCCCAGGGTGCCGGGCCGATCGCCCGGGTCGGAGGCGGTG

41761 + + + + + + 41820

AGCGGGTAGTCCTCGTCCACCGGGTCCCACGGCCCGGCTAGCGGGCCCAGCCTCCGCCAC
EGMLLLHGLTGPRDGPDSAT-

ATCACTCCCAGGAGGTCCCGGGCGAAGCTGGGCACGGTCTGGAGATGAGTGATCCGCTCC

41821 + + + + + + 41880

TAGTGAGGGTCCTCCAGGGCCCGCTTCGACCCGTGCCAGACCTCTACTCACTAGGCGAGG
IVGLLDRAFSPVTQLHTIRE-

TGGACGAGCCACGGCACCAGCTTGTCGGGGTTCACCCTGACGCGCTCCGGCACCGGACAC

41881 + + + + + + 41940

ACCTGCTCGGTGCCGTGGTCGAACAGCCCCAAGTGGGACTGCGCGAGGCCGTGGCCTGTG

QVLWPVLKDPNVRVREPVPC-

AGCGTCCCGCCGGCCACGAGCGTCGCGAAGACCTCGGCCAGCGCCGGGTCGTGCTCCGGG

41 941 + + + + + + 42000

TCGCAGGGCGGCCGGTGCTCGCAGCGCTTCTGGAGCCGGTCGCGGCCCAGCACGAGGCCC
LTGGAVLTAFVEALAPDHEP -

SEQ ID No. 2. C-1027 gene cluster DNA sequence from 41,980 to 63,164

AGCGCCGGGTCGTGCTCCGGGGAGACCCACTGCGCCACCCGCGCGCCCGGCCCCATCGCG

41980 + + + + + + 42039

TCGCGGCCCAGCACGAGGCCCCTCTGGGTGACGCGGTGGGCGCGCGGGCCGGGGTAGCGC

LAPDHEPSVWQAVRAGPGMA-

AACCGTTCGCCCATCCAGCCCGCGAACTGGCCCAGCGCGGCATGCGACTGGGCGATCCCC

42040 + + + + + + 42099

TTGGCAAGCGGGTAGGTCGGGCGCTTGACCGGGTCGCGCCGTACGCTGACCCGCTAGGGG
FREGMWGAFQGLAAHSQAIG-

TTGGGCCGCCCGGTCGAACCCGAGGTGAACGCCACGTAGGCCAGGTCTGCCAGGCCCGGC

42100 + + + + + + 42159

AACCCGGCGGGCCAGCTTGGGCTCCACTTGCGGTGCATCCGGTCCAGACGGTCCGGGCCG

53

25 KPRGTSGSTFAVYALDALGP-

CCCGCCGCGGTCGTCGCGTCCGGGCCGGCGGCGGGTCGAGGGCCGAGCACAGAGGAGGCG

42160 + + + + + + 42219

GGGCGGCGCCAGCAGCGCAGGCCCGGCCGCCGCCCAGCTCCCGGCTCGTGTCTCCTCCGC
25 GAATTADPGAAPRPGLVSSA-

TCCAGCAGGGTGGCGCCCGGTTCACCGGCGTACCAGAGCGCCAGCGGATCCTCCTGCGGA

42220 + + + + + + 42279

AGGTCGTCCCACCGCGGGCCAAGTGGCCGCATGGTCTCGCGGTCGCCTAGGAGGACGCCT
25 DLLTAGPEGAYWLALPDEQP

TCGCCGTCGAGGACCAGGCACGCCGGGCGCAGATCGCTGAGCATCGACCGGTGTCGTTCG

42280 + + + + + + 42339

AGCGGCAGCTCCTGGTCCGTGCGGCCCGCGTCTAGCGACTCGTAGCTGGCCACAGCAAGC
25 DGDLVLCAPRLDSLMSRHRE-

CCCGCGCCGTCCGGAGCGAACCACGCCAGGTGGGCGCCCGCCTCCAGGACTCCCAGCAGC

42340 + + + + + + 42399

GGGCGCGGCAGGCCTCGCTTGGTGCGGTCCACCCGCGGGCGGAGGTCCTGAGGGTCGTCG
25 GAGDPAFWALHAGAELVGLL-

ACCGCGATCCGGCGGGCGCCCGGCTGCATCCGCACCGCCACCGGCGAGCCGTGCCCCGCG

42400 + + + + + + 42459

TGGCGCTAGGCCGCCCGCGGGCCGACGTAGGCGTGGCGGTGGCCGCTCGGCACGGGGCGC
25 VAIRRAGPQMRVAVPSGHGA-

CCGGCCGCGGTGAGGGCCGAGGCGACGCGGGCCGCGTCCGCGGTCAGTTCGGCGGTCAGT

42460 + + + + + + 42519

GGCCGGCGCCACTCCCGGCTCCGCTGCGCCCGGCGCAGGCGCCAGTCAAGGCGCCAGTCA
25 GAATLASAVRAADATLEATL

TCGGCGTAGCTTGTGCGCGTGCCGCCGAACGAGACGGCGACACCGTCGTGTTCCGCGTGG

42520 + + + + + + 42579

AGCCGCATCGAACACGCGCACGGCGGCTTGCTCTGCCGCTGTGGCAGCACAAGGCGCACC
25 EAYSTRTGGFSVAVGDHEAH

CGGCGGACCGAGGCGTGCACCGGCCGCGTCATGTCCCCGCCGGACGCCCGGCGGTCCGAA

42580 + + + + + + 42639

GCCGCCTGGCTCCGCACGTGGCCGGCGCAGTACAGGGGCGGCCTGCGGGCCGCCAGGCTT
25 RRVSAHVPRTM (ORF25)

GCGCGCAGGGCGTGGTCCCGGTGGCGGTCGTCGTCCAGCGGCAGAGCGCCCACGGGTGTG

42640 + + + + + + 42699

CGCGCGTCCCGCACCAGGGCCACCGCCAGCAGCAGGTCGCCGTCTCGCGGGTGCCCACAC

TCCGGATCCGTGGTCGCGGCGGTCAGGAGGACGGCCAGCTGATCCAGCATCCGCCGGGCC

42700 + + + + + + 42759

AGGCCTAGGCACCAGCGCCGCCAGTCCTCCTGCCGGTCGACTAGGTCGTAGGCGGCCCGG

GAAGCGGGCTCGAACAGAGCTTCGCGGTACTCCAGGTAGCCGGTGACCGAGGGCGCGGTG

42760 + + + + + + 42819

CTTCGCCCGAGCTTGTCTCGAAGCGCCATGAGGTCCATCGGCCACTGGCTCCCGCGCCAC

TCCTGCAGCACCAGGGTCAGGTCGGCGGCGGCAGTGCCGTTGTGCACGGACAGCCGCCTC

42820 + + + + + + 42879

AGGACGTCGTGGTCCCAGTCCAGCCGCCGCCGTCACGGCAACACGTGCCTGTCGGCGGAG

ACCTCGGCGCCTGGTATCCGCAGGCCCGGCCGCTCCTCGTGGACGAACACGGCGTCGGCC

42880 + + + + + + 42939

TGGAGCCGCGGACCATAGGCGTCCGGGCCGGCGAGGAGCACCTGCTTGTGCCGCAGCCGG

CCCTCGATCCGGCACGGCCCGGGGGCCGGGGCCGGCGTCGTGTGCAGCAGCTCCCGGAAG

42940 + + + + + + 42999

GGGAGCTAGGCCGTGCCGGGCCCCCGGCCCCGGCCGCAGCACACGTCGTCGAGGGCCTTC

54

GCGGTGGCCGGCGTGCCGTCGTCCTGTCCGGCGTAGCGCTGGACCAGGGCTCGGAATCCG

43000 + + + + + + 43059

CGCCACCGGCCGCACGGCAGCAGGACAGGCCGCATCGCGACCTGGTCCCGAGCCTTAGGC

GCCAGCACCACGGCCGCGGCGGTGACCCCTTCCGCTTCGGCGAGCCGGGCCGTACGGAAG

43060 + + + + + + 43119

CGGTCGTGGTGCCGGCGCCGCCACTGGGGAAGGCGAAGCCGCTCGGCCCGGCATGCCTTC

CCGAGGTCCGGACTCCAGCCGAAGGCGACGGTGCTCCCCGCGTGCGAGGGCAGGTGCGGG

43120 + + + + + + 43179

GGCTCCAGGCCTGAGGTCGGCTTCCGCTGCCACGAGGGGCGCACGCTCCCGTCCACGCCC

CGGTTCCGGTCGGCGGGCAGGACCTGTCCGGAGGCGGTCGCCGAAGACTCCTCGCTCCCG

43180 + + + + + + 43239

GCCAAGGCCAGCCGCCCGTCCTGGACAGGCCTCCGCCAGCGGCTTCTGAGGAGCGAGGGC

GGCGCCCGGGGCGTTTGCGGCGCGGGCGCAGTGGGAGGCCGGCCGCCGGTGGTGACGGCG

43240 + + + + + + 43299

CCGCGGGCCCCGCAAACGCCGCGCCCGCGTCACCCTCCGGCCGGCGGCCACCACTGCCGC

AGGTACGCGTTCGACAACGCGGCCGGCAGGGGCCCGGACGGCCCGTCCCAGGCTCCGGAG

43300 + + + + + + 43359

TCCATGCGCAAGCTGTTGCGCCGGCCGTCCCCGGGCCTGCCGGGCAGGGTCCGAGGCCTC

TGCGAGGCCACCAGGAGAAGCAGGTGCGCGCGTGGGCCTCTGCGGGCGATGTGGAGCCGT

43360 + + + + + + 43419

ACGCTCCGGTGGTCCTCTTCGTCCACGCGCGCACCCGGAGACGCCCGCTACACCTCGGCA

GCGGGCGCGTCACCCTCGGCGAAGGGACGGGCCGCCCAGCGAGCGCAGAGTTCCTCCTCC

43420 + + + + + + 43479

CGCCCGCGCAGTGGGAGCCGCTTCCCTGCCCGGCGGGTCGCTCGCGTCTCAAGGAGGAGG

CCGCACTCCTCGTCGGCACTCGGCCCGTCCACGGCGGCCCCGTCTCCGGCGGCGGCCCGC

43480 + + + + + + 43539

GGCGTGAGGAGCAGCCGTGAGCCGGGCAGGTGCCGCCGGGGCAGAGGCCGCCGCCGGGCG

CAGGCCGTCCGCAGGGCCTCCAGGTCGAGTCCGCCGCTCACGTGGTAGGCCGCGTACGGG

43540 + + + + + + 43599

GTCCGGCAGGCGTCCCGGAGGTCCAGCTCAGGCGGCGAGTGCACCATCCGGCGCATGCCC

TGCAACACCGCAGATCCGGAGGCCGGCGAAGGCCCCCGGTCCGGCTCGGTCACAGTCACG

43600 + + + + + + 43659

ACGTTGTGGCGTCTAGGCCTCCGGCCGCTTCCGGGGGCCAGGCCGAGCCAGTGTCAGTGC

TCATTCGCCACGACGCCCATCTTGGGGCGGCGGCGCACAGGACGCTTCTCCTTGAGTGCG

43660 + + + + + + 43719

AGTAAGCGGTGCTGCGGGTAGAACCCCGCCGCCGCGTGTCCTGCGAAGAGGAACTCACGC

GAGCTCCGCGTACGGCGCCGAAGCGTTCGGTCAAACCTTGTTCGACCAACTGCGCAATCT

43720 + + + + + + 43779

CTCGAGGCGCATGCCGCGGCTTCGCAAGCCAGTTTGGAACAAGCTGGTTGACGCGTTAGA

GGAAGTTGACGTCTTCCAGGTGGAGTTGGGAACGATGGAGGCCCCCGCCGGCCGCGTCGG

43780 + + + + + + 43839

CCTTCAACTGCAGAAGGTCCACCTCAACCCTTGCTACCTCCGGGGGCGGCCGGCGCAGCC

AACGGCCGTGCAGTGCGGCCCTCTCCAACACTCCCGGCCATCGCGGAATCCGAGACGTGC

43840 + + + + + + 43899

TTGCCGGCACGTCACGCCGGGAGAGGTTGTGAGGGCCGGTAGCGCCTTAGGCTCTGCACG

CCGAAGGAGCCCCCCTTGCAAGCCTGGTTCAAGCGCACCAGTGGTGTGCCCGGTGACAGA

43900 + + + + + + 43959

GGCTTCCTCGGGGGGAACGTTCGGACCAAGTTCGCGTGGTCACCACACGGGCCACTGTCT
27 (ORF27) V P G D R

55

CGTGGAAAGTGGCTGGTCCTGGCCGCCTGGCTCATCATCGCGATGGCGCTGGGCCCGCTG

43960 + + + + + + 44019

GCACCTTTCACCGACCAGGACCGGCGGACCGAGTAGTAGCGCTACCGCGACCCGGGCGAC
27 RGKWLVLAAWLI IAMALGPL

GCGGGGAAGCTCGCCGACGTCCAGGACTCCAGCGCCAACGCCTTCCTTCCGCGCAGCTCG

44020 + + + + + + 44079

CGCCCCTTCGAGCGGCTGCAGGTCCTGAGGTCGCGGTTGCGGAAGGAAGGCGCGTCGAGC
27 AGKLADVQDS SANAFLPRS S

GAGTCCGCGAAGCTGAACAAGGAACTGGAGAAGTTCCGCGCCGACGAGCTGATGCCGGCC

44080 + + + + + + 44139

CTCAGGCGCTTCGACTTGTTCCTTGACCTCTTCAAGGCGCGGCTGCTCGACTACGGCCGG
27 ESAKLNKELEKFRADELMPA

GTGGTGGTCTACAGCGCCGACGGCTCGCTGCCCGCCGAGGGGCGGGCCAAGGCCGAGAAG

44140 + + + + + + 44199

CACCACCAGATGTCGCGGCTGCCGAGCGACGGGCGGCTCCCCGCCCGGTTCCGGCTCTTC
27 VVVYSADGSLPAEGRAKAEK

GACATAGC CGCCTTCCAGGAGCTGGCCGC CGAGGGCGAGAAGGTCGAAGCGCCCCTGGAG

44200 + + + + + + 44259

CTGTATCGGCGGAAGGTCCTCGACCGGCGGCTCCCGCTCTTCCAGCTTCGCGGGGACCTC
27 DIAAFQELAAEGEKVEAPLE

TCGGAGGACGGCCAGGCGCTCATGGTCGTCGTTCCGCTGATCAGCGACGCCGACATCGTC

44260 + + + + + + 44319

AGCCTCCTGCCGGTCCGCGAGTACCAGCAGCAAGGCGACTAGTCGCTGCGGCTGTAGCAG
27 SEDGQALMVVVPLI SDADIV

GCCACGACGAAGAAGGTCCGCGATGTCGCGGACGCCAACGCCCCCCCGGGCGTCGCCATC

44320 + + + + + + 44379

CGGTGCTGCTTCTTCCAGGCGCTACAGCGCCTGCGGTTGCGGGGGGGCCCGCAGCGGTAG
27 ATTKKVRDVADANAPPGVAI

GAGGTGGGCGGGCCCGCCGGGTCGACGACCGACGCCGCCGGCGCTTTCGAGTCCCTCGAC

44380 + + + + + + 44439

CTCCACCCGCCCGGGCGGCCCAGCTGCTGGCTGCGGCGGCCGCGAAAGCTCAGGGAGCTG
27 EVGGPAGSTTDAAGAFESLD

TCCATGCTGATGATGGTCACCGGCCTTGTGGTCGCCATCCTGCTGCTGATCACCTACCGC

44440 + + + + + + 44499

AGGTACGACTACTACCAGTGGCCGGAACACCAGCGGTAGGACGACGACTAGTGGATGGCG
27 SMLMMVTGLVVAILLLITYR

TCCCCCATCCTGTGGCTGCTGCCCCTGCTCTCCGTCGGCTTCGCCTCCGTGCTGACCCAG

44500 + + + + + + 44559

AGGGGGTAGGACACCGACGACGGGGACGAGAGGCAGCCGAAGCGGAGGCACGACTGGGTC
27 SPILWLLPLLSVGFASVLTQ

GTCGGCACCTACATGCTCGCCAAGTACGCCGGGCTGCCGGTCGACCCGCAGAGCTCCGGC

44560 + + + + + + 44619

CAGCCGTGGATGTACGAGCGGTTCATGCGGCCCGACGGCCAGCTGGGCGTCTCGAGGCCG
27 VGTYMLAKYAGLPVDPQSSG

GTCCTGATGGTCCTCGTGTTCGGTGTCGGCACCGACTACGCCCTGCTGCTCATCGCCCGC

44620 + + + + + + 44679

CAGGACTACCAGGAGCACAAGCCACAGCCGTGGCTGATGCGGGACGACGAGTAGCGGGCG
27 VLMVLVFGVGTDYALLLIAR

TACCGTGAGGAACTGCGCCGCGAGCAGGACCGGCACGTGGCCATGAAGACCGCGTTGCGA

44680 + + + + + + 44739

ATGGCACTCCTTGACGCGGCGCTCGTCCTGGCCGTGCACCGGTACTTCTGGCGCAACGCT
27 YREELRREQDRHVAMKTALR

CGGTCGGGCCCGGCCATCCTGGCCTCGGCCGGCACCATCGCCATCGGCCTCGTCTGCCTG

56

44740 + + + + + + 44799

GCCAGCCCGGGCCGGTAGGACCGGAGCCGGCCGTGGTAGCGGTAGCCGGAGCAGACGGAC
27 RSGPAILASAGTIAIGLVCL

GTCCTCGCGGACGTCAACTCCTCCCGCTCCATGGGCCTGGTCGGCGCGATCGGCGTGGTC

44800 + + + + + + 44859

CAGGAGCGCCTGCAGTTGAGGAGGGCGAGGTACCCGGACCAGCCGCGCTAGCCGCAGCAG
27 VLADVNS SRSMGLVGAI GVV

TGCGCCCTCCTCGCCATGGTCACGATCCTGCCCGCGCTGCTGGTCATCCTGGGCCGCTGG

44860 + + + + + + 44919

ACGCGGGAGGAGCGGTACCAGTGCTAGGACGGGCGCGACGACCAGTAGGACCCGGCGACC
27 CALLAMVTILPALLVILGRW

GTGTTCTGGCCCTTCGTTCCCCGCTGGACGCCGGAGTCGGCCGCGGCCCCCGAGGCACCG

44920 + + + + + + 44979

CACAAGACCGGGAAGCAAGGGGCGACCTGCGGCCTCAGCCGGCGCCGGGGGCTCCGTGGC
27 VFWPFVPRWTPESAAAPEAP

GCGTCCCACAGCCGCTGGGAGCGCATCGGCTCCGTCACGGCCGCCCGGCCGCGCCGCGCC

44980 + + + + + + 45039

CGCAGGGTGTCGGCGACCCTCGCGTAGCCGAGGCAGTGCCGGCGGGCCGGCGCGGCGCGG
27 ASHSRWERIGSVTAARPRRA

TGGGTGCTGTCCTTGGCCGCGACGGGGCTTCTCGCCCTCAGTTCCCTCGGCCTCGACATG

45040 + + + + + + 45099

ACCCACGACAGGAACCGGCGCTGCCCCGAAGAGCGGGAGTCAAGGGAGCCGGAGCTGTAC
27 WVLSLAATGLLALS S LGLDM

GGACTCACCCAGAGCGAACTGCTCCAGACGAAGCCCGAGTCCGTCGTCGCCCAGGAGCGG

45100 + + + + + + 45159

CCTGAGTGGGTCTCGCTTGACGAGGTCTGCTTCGGGCTCAGGCAGCAGCGGGTCCTCGCC
27 GLTQSELLQTKPESVVAQER

ATCTCCGCCCACTACCCGTCCGGCTCCTCCGACCCCGCCACCGTCGTCGCACCCAGCGCG

45160 + + + + + + 45219

TAGAGGCGGGTGATGGGCAGGCCGAGGAGGCTGGGGCGGTGGCAGCAGCGTGGGTCGCGC
27 ISAHYPSGSSDPATVVAPSA

GACGTGGCCGAGGTCCGCCGGGCCGCCGAGGGGACCGACGGAGTGGTCTCCGTCCAGGAC

45220 + + + + + + 45279

CTGCACCGGCTCCAGGCGGCCCGGCGGCTCCCCTGGCTGCCTCACCAGAGGCAGGTCCTG
27 DVAEVRRAAEGTDGVVSVQD

GGCCCCACCACTCCCGACGGAGAGCTGACCATGCTGTCCGTGGTGCTGAAGGACGTTCCC

45280 + + + + + + 45339

CCGGGGTGGTGAGGGCTGCCTCTCGACTGGTACGACAGGCACCACGACTTCCTGCAAGGG
27 GPTTPDGELTMLSVVLKDVP

GACAGCAGCGGGGCCAAGGACACCATCGATGCACTGCGGGACAACACGGATGCTCTCGTG

45340 + + + + + + 45399

CTGTCGTCGCCCCGGTTCCTGTGGTAGCTACGTGACGCCCTGTTGTGCCTACGAGAGCAC
27 DSSGAKDTIDALRDNTDALV

GGGGGTACGACGGCCCAGAGCCTGGACACCCAGCGCGCCTCGGTCCGTGACCTCTGGGTC

45400 + + + + + + 45459

CCCCCATGCTGCCGGGTCTCGGACCTGTGGGTCGCGCGGAGCCAGGCACTGGAGACCCAG
27 GGTTAQSLDTQRASVRDLWV

ACCGTCCCCGCGGTCCTGCTGGTGGTCCTGCTCGTCCTGATCTGGCTGCTGCGCTCGGTC

45460 + + + + + + 45519

TGGCAGGGGCGCCAGGACGAC CAC CAGGACGAGCAGGACTAGAC CGACGACGCGAGCCAG
27 TVPAVLLVVLLVLIWLLRSV

ACCGGACCGCTGATCATGCTCGGCACCGTGGTCGTGTCGTTCTTCGCGGCCCTGGGGGCG
45520 + + + + + + 45579

57

TGGCCTGGCGACTAGTACGAGCCGTGGCAC CAGCACAGCAAGAAGCGC CGGGACCCCCGC
27 TGPL I MLGTVVVS FFAALGA

TCCAACCTGCTCTTCGAGTACGTGATGGGGCACGCCGGCGTCGACTGGTCGGTGCCGCTT

45580 + + + + + + 45639

AGGTTGGACGAGAAGCTCATGCACTACCCCGTGCGGCCGCAGCTGACCAGCCACGGCGAA

27 SNLLFEYVMGHAGVDWSVPL

CTCGGGTTCGTGTACCTGGTCGCCCTCGGAATCGACTACAACATCTTCCTCATGCACCGG

45640 + + + + + + 45699

GAGCCCAAGCACATGGACCAGCGGGAGCCTTAGCTGATGTTGTAGAAGGAGTACGTGGCC
27 LGFVYLVALGIDYNI FLMHR

GTGAAGGAGGAGGTCGCTCTGCACGGCCATGCCAAGGGCGTGCTCACCGGCCTGACCACC

45700 + + + + + + "-- 45759

CACTTCCTCCTCCAGCGAGACGTGCCGGTACGGTTCCCGCACGAGTGGCCGGACTGGTGG
27 VKEEVALHGHAKGVLTGLTT

ACCGGGGGCGTCATCACCAGTGCCGGCGTGGTCCTGGCCGCGACGTTCGCCGTCATCGCC

45760 + + + + + + 45819

TGGCCCCCGCAGTAGTGGTCACGGCCGCACCAGGACCGGCGCTGCAAGCGGCAGTAGCGG

27 TGGVI TSAGVVLAATFAVIA

ACACTGCCGCTGGTCCCGATGGCCCAGATGGGTGTCGTGGTCGGCCTGGGCATTCTGCTG

45820 + + + + + + 45879

TGTGACGGCGACCAGGGCTACCGGGTCTACC CACAGCACCAGCCGGACC CGTAAGACGAC
27 TLPLVPMAQMGVVVGLGILL

GACACCTTCCTCGTCCGGACGATTCTTCTGCCGGCCCTGGCGCTCGATCTGGGGCCCCGG

45880 + + + + + + 45939

CTGTGGAAGGAGCAGGCCTGCTAAGAAGACGGCCGGGACCGCGAGCTAGACCCCGGGGCC
27 DTFLVRTILLPALALDLGPR

TTCTGGTGGCCGGGCGCGCTGTCGAAGACGTCCGGGGGACCGGCCCCCGTCCGCGAGGAC

45940 + + + + + + 45999

AAGACCACCGGCCCGCGCGACAGCTTCTGCAGGCCCCCTGGCCGGGGGCAGGCGCTCCTG
27 FWWPGALSKTSGGPAPVRED

CGCACGTCCCAGCCCGTGGGCTGAGACCCGTCCCGACGAGACCCGTACGGCGGGCGGCCG

46000 + + + + + + 46059

GCGTGCAGGGTCGGGCACCCGACTCTGGGCAGGGCTGCTCTGGGCATGCCGCCCGCCGGC

27 RTSQPVG* (ORF27)

GTTCCCCCGGGCCGTACGACTGAGCAACCCAGAAGATGGGCCGCCCGCGACCAGGCGTCA

46060 + + + + + + 46119

CAAGGGGGCCCGGCATGCTGACTCGTTGGGTCTTCTACCCGGCGGGCGCTGGTCCGCAGT

CGATGGTGGCCCACCGGCCGCAGGCCGATCTCCCGGAAGGAAGCGCCGTGTTGGGCGATG

46120 + + + + + + 46179

GCTACCACCGGGTGGCCGGCGTCCGGCTAGAGGGCCTTCCTTCGCGGCACAACCCGCTAC

28 (ORF2 8) V L G D E -

AGGACGGCAAGGCCGCCGAGCTGTGGTCGATGGCGAACCTGGGTACACCGATGGCCGTGC

46180 + + + + + + 4 &239

TCCTGCCGTTCCGGCGGCTCGACACCAGCTACCGCTTGGACCCATGTGGCTACCGGCACG

28 DGKAAELWSMANLGTPMAVR-

GCGTCGCGGCGACCCTGCGCATCGCCGACCACATCACGGCCGGAGCGCACACCGCCGGCG

46240 + + + + + + 4 "99

CGCAGCGCCGCTGGGACGCGTAGCGGCTGGTGTAGTGCCGGCCTCGCGTGTGGCGGCCGC
28 VAATLRIADHI TAGAHTAGE-

AAATCGCCGAAGCGGCCGCCGTGCACGAGGAATCCCTCGACCGGCTGCTGCGCTACCTCA

46300 + + + + + + 46359

TTTAGCGGCTTCGCCGGCGGCACGTGCTCCTTAGGGAGCTGGCCGACGACGCGATGGAGT
28 IAEAAAVHEESLDRLLRYLT-

58

CCGTCCGGGGCCTGCTGGACCGTGACGGGCTCGGCCGGTACACGCTGACCCCCCTGGGCC

46360 + + + + + + 46419

GGCAGGCCCCGGACGACCTGGCACTGCCCGAGCCGGCCATGTGCGACTGGGGGGACCCGG
28 VRGLLDRDGLGRYTLTPLGR-

GGCCGCTGTGCGAGGACCACCCCGCCGGCGTCCGGGCCTGGTTCGACATGGAGGGAGCGG

46420 + + + + + + 46479

CCGGCGACACGCTCCTGGTGGGGCGGCCGCAGGCCCGGACCAAGCTGTACCTCCCTCGCC
28 PLCEDHPAGVRAWFDMEGAG-

GGCGGGGCGAGCTGTCGTTCGTCGACCTGCTGCACAGCGTACGGACCGGGAAGGCCGCCT

46480 + + + + + + 46539

CCGCCCCGCTCGACAGCAAGCAGCTGGACGACGTGTCGCATGCCTGGCCCTTCCGGCGGA
28 RGELSFVDLLHSVRTGKAAF-

TCCCCCTGCGCTACGGCCGCCCCTTCTGGGAGGACCTGGCGGAGGACCCCCGCCGCGCGG

46540 + + + + + + 46599

AGGGGGACGCGATGCCGGCGGGGAAGACCCTCCTGGACCGCCTCCTGGGGGCGGCGCGCC
28 PLRYGRPFWEDLAED PRRAE-

AGTCCTTCAACCGGCTGCTCGGCCAGGACGTCGCCACTCGCGCCCCGGCCGTGGTGGCCG

46600 + + + + + + 46659

TCAGGAAGTTGGCCGACGAGCCGGTCCTGCAGCGGTGAGCGCGGGGCCGGCACCACCGGC
28 SFNRLLGQDVATRAPAVVAG-

GCTTCGACTGGGCGAGCACCGGTCATGTCATCGACCTCGGAGGCGGCGACGGCTCCCTGC

46660 + + + + + + 46719

CGAAGCTGACCCGCTCGTGGCCAGTACAGTAGCTGGAGCCTCCGCCGCTGCCGAGGGACG
28 FDWASTGHVIDLGGGDGSLL-

TGACCGCACTGCTGACCGCCTGTCCGTCACTGCGCGGCACGGTCCTGGACCTGCCCGAAG

46720 + + + + + + 4 6 77 9

ACTGGCGTGACGACTGGCGGACAGGCAGTGACGCGCCGTGCCAGGACCTGGACGGGCTTC
28 TALLTAC P SLRGTVLDL P EA-

CGGTGCAGCGTGCCAAGGAGTCGTTCGCCGTGTCCGGACTGGACGACCGGGCGAACGCGG

46780 + + + + + + 46839

GCCACGTCGCACGGTTCCTCAGCAAGCGGCACAGGCCTGACCTGCTGGCCCGCTTGCGCC
28 VQRAKES FAVSGLDDRANAV-

TCGCGGGCAGCTTCTTCGACGCCCTCCCCGCCGGCGCGGGCGCCTACGTCCTGTCCCTGG

46840 + + + + + + 46899

AGCGCCCGTCGAAGAAGCTGCGGGAGGGGCGGCCGCGCCCGCGGATGCAGGACAGGGACC
28 AGS FFDALPAGAGAYVLSLV-

TCCTGCACGACTGGGACGACGAGGCGTCCGTCGCGATCCTGCGGCGCTGCGCCGAGGCGG

46900 + + + + + + 46959

AGGACGTGCTGACCCTGCTGCTCCGCAGGCAGCGCTAGGACGCCGCGACGCGGCTCCGCC
28 LHDWDDEASVAI LRRCAEAA-

CGGGGCAGACGGGATCGGTGTTCGTCATCGAGTCGACCGGCTCGGCGGGGGACGCCCCGC

46960 + + + + + + 4 7019

GCCCCGTCTGCCCTAGCCACAAGCAGTAGCTCAGCTGGCCGAGCCGCCCCCTGCGGGGCG
28 GQTGSVFVI ESTGSAGDAPH-

ACACAGGTATGGACCTGCGCATGCTGTGCATCTACGGAGCCAAGGAGCGCCGCGTGGAGG

47020 + + + + + + 47079

TGTGTCCATACCTGGACGCGTACGACACGTAGATGCCTCGGTTCCTCGCGGCGCACCTCC
28 TGMDLRMLCIYGAKERRVEE-

AGTTCGAGGAACTCGCCGGCCGGGCCGGGCTCCGGGTCGTCGCCGTCCACCCCGCGGGCC

47080 + + + + + + 47139

TCAAGCTCCTTGAGCGGCCGGCCCGGCCCGAGGCCCAGCAGCGGCAGGTGGGGCGCCCGG
28 FEELAGRAGLRVVAVH PAGP-

59

CTTCCGCGATCATCCAGATGTCCGCGGTCTGACCGCCCGGAGCCCCGGCCCATCGCGGCG

47140 + + + + + + 47199

GAAGGCGCTAGTAGGTCTACAGGCGCCAGACTGGCGGGCCTCGGGGCCGGGTAGCGCCGC

28 SAIIQMSAV* (ORF28)

CGGGCCACGGCAGACAAGGAGAGAGCGTATGGCCGGCCTGGTCATGTCGCCGGTGGAGGC
47200 + + + + + + 47259

GCCCGGTGCCGTCTGTTCCTCTCTCGCATACCGGCCGGACCAGTACAGCGGCCACCTCCG

(ORF29) MAGLVMS P V E A -

GCTCGACGCGCTGGGCACGGTGCAGGGGCGTCAGGACCCCTATCCCTTCTACGAGGCGAT

47260 + + + + + + 47319

CGAGCTGCGCGACCCGTGCCACGTCCCCGCAGTCCTGGGGATAGGGAAGATGCTCCGCTA
29 LDALGTVQGRQDPYPFYEAI

CCGCGCGCACGGGCAGGCGGTCCCCACGAAGCCCGGCCGCTTCGTGGTGGTCGGCCACGA
47320 + + + + + + 47379

GGCGCGCGTGCCCGTCCGCCAGGGGTGCTTCGGGCCGGCGAAGCACCACCAGCCGGTGCT

29 RAHGQAVPTKPGRFVVVGHD

CGCGTGCGACCGGGCGCTGCGGGAACCGGCCCTGCGCGTCCAGGACGCCAGGAGCTACGA

47380 + + + + + + 47439

GCGCACGCTGGCCCGCGACGCCCTTGGCCGGGACGCGCAGGTCCTGCGGTCCTCGATGCT
29 ACDRALREPALRVQDARSYD

CGTCGTCTTCCCCTCGTGGCGGTCGCACTCCTCGGTCCGGGGGTTCACCAGCTCCATGCT

47440 + + + + + + 47499

GCAGCAGAAGGGGAGCACCGCCAGCGTGAGGAGCCAGGCCCCCAAGTGGTCGAGGTACGA
29 VVFPSWRSHSSVRGFTSSML-

CTACAGCAACCCGCCCGATCACGGCCGGTTGCGCCAGGTGGTGAGCTTCGCGTTCACCCC

47500 + + + + + + 47559

GATGTCGTTGGGCGGGCTAGTGCCGGCCAACGCGGTCCACCACTCGAAGCGCAAGTGGGG
29 YSNPPDHGRLRQVVSFAFTP

GCCCAAGGTGCGCCGGATGCACGGGGTGATCGAGGACATGACCGACCGGCTCCTCGACCG
47560 + + + + + + 47619

CGGGTTCCACGCGGCCTACGTGCCCCACTAGCTCCTGTACTGGCTGGCCGAGGAGCTGGC
29 PKVRRMHGVIEDMTDRLLDR

GATGGCCCGGCTCGGCTCCGGCGGCTCCCCGGTCGACCTCATAGCCGAGTTCGCCGCCCG
47620 + + + + + + 47679

CTACCGGGCCGAGCCGAGGCCGCCGAGGGGCCAGCTGGAGTATCGGCTCAAGCGGCGGGC
29 MARLGSGGSPVDLIAEFAAR-

GCTGCCCGTCGCGGTGATCAGCGAGATGATCGGCTTTCCGGCGAAGGACCAGGTGTGGTT

47680 + , + + + + + 47739

CGACGGGCAGCGCCACTAGTCGCTCTACTAGCCGAAAGGCCGCTTCCTGGTCCACACCAA
29 LPVAVISEMIGFPAKDQVWF

CCGCGACATGGCCTCCCGGGTCGCCGTGGCGACGGACGGTTTCACCGACCCCGGCGCGCT

47740 + + + + + + 47799

GGCGCTGTACCGGAGGGCCCAGCGGCACCGCTGCCTGCCAAAGTGGCTGGGGCCGCGCGA
29 RDMASRVAVATDGFTDPGAL

CACGGGGGCCGACGCCGCCATGGACGAGATGAGCGCCTACTTCGACGACCTCCTGGACCG

47800 + + + + + + 47859

GTGCCCCCGGCTGCGGCGGTACCTGCTCTACTCGCGGATGAAGCTGCTGGAGGACCTGGC
29 TGADAAMDEMSAYFDDLLDR

TCGCCGCCGCACCCCGGCCGACGACCTGGTCACCCTGCTCGCCGAGGCCCACGACGGCTC
47860 + + + + + + 47919

AGCGGCGGCGTGGGGCCGGCTGCTGGACCAGTGGGACGAGCGGCTCCGGGTGCTGCCGAG
29 RRRTPADDLVTLLAEAHDGS

CCCCGGGCGCCTGGACCACGACGAACTGATGGGCACCATGATGGTGCTGCTCACAGCCGG

60

47920 + + + + + + 47979

GGGGCCCGCGGACCTGGTGCTGCTTGACTACCCGTGGTACTACCACGACGAGTGTCGGCC
29 PGRLDHDELMGTMMVLLTAG-

GTTCGAGACCACGAGCTTTCTGATCGGCCACGGGGCGATGATCGCCCTCGAACAACGGGC

47980 + + + + + + 48039

CAAGCTCTGGTGCTCGAAAGACTAGCCGGTGCCCCGCTACTAGCGGGAGCTTGTTGCCCG
29 FETTSFLIGHGAMIALEQRA-

GCACGCGGCCCGGCTGCGGGCCGAACCCGACTTCGCCGACGGCTACGTCGAGGAGATCCT

48040 + + + + + + 48099

CGTGCGCCGGGCCGACGCCCGGCTTGGGCTGAAGCGGCTGCCGATGCAGCTCCTCTAGGA
29 HAARLRAEPDFADGYVEE I L

CAGGTTCGAGCCGCCGGTCCACGTCACCAGCCGGTGGGCTGCCGAGGACCTCGACCTGCT

48100 + + + + + + 48159

GTCCAAGCTCGGCGGCCAGGTGCAGTGGTCGGCCACCCGACGGCTCCTGGAGCTGGACGA
29 RFEPPVHVTSRWAAEDLDLL-

GGGCCTGTCCGTACCGGCGGGCTCCAAGCTGGTCCTGATCCTGGCCGCCGCGAATCGCGA

48160 + + + + + + 48219

CCCGGACAGGCATGGCCGCCCGAGGTTCGACCAGGACTAGGACCGGCGGCGCTTAGCGCT
29 GLSVPAGSKLVLI LAAANRD

TCCCGGCCGCTACCCCGAGCCCGGCCGCTTCGACCCCGACCGCTACGCGCCCCGGCCGGG

48220 + + + + + + 48279

AGGGCCGGCGATGGGGCTCGGGCCGGCGAAGCTGGGGCTGGCGATGCGCGGGGCCGGCCC
29 PGRYPEPGRFDPDRYAPRPG-

CGGGCCGGAGGCCACCAGACCGCTGAGCTTCGGCGCGGGCGGCCACTTCTGCCTCGGCGC

48280 + + + + + + 48339

GCCCGGCCTCCGGTGGTCTGGCGACTCGAAGCCGCGCCCGCCGGTGAAGACGGAGCCGCG
29 GPEATRPLSFGAGGHFCLGA-

TCCGCTGGCGCGGCTGGAAGCCCGGATCGCGCTGCCGCGTCTGCTGCGCCGCTTCCCGGA

48340 + + + + + + 48399

AGGCGACCGCGCCGACCTTCGGGCCTAGCGCGACGGCGCAGACGACGCGGCGAAGGGCCT
29 PLARLEARIALPRLLRRFPD-

CCTGGCCGTGTCCGAGCCCCCCGTCTACCGCGACCGCTGGGTCGTCCGCGGCCTCGAAAC

48400 + + + + + + 48459

GGACCGGCACAGGCTCGGGGGGCAGATGGCGCTGGCGACCCAGCAGGCGCCGGAGCTTTG
29 LAVSEPPVYRDRWVVRGLET-

CTTTCCCGTGACCCTCGGGTCCTGAGCCCCCGCCGGCCGGAACACGTGACCGTCCCGGCC

48460 + + + + + + 48519

GAAAGGGCACTGGGAGCCCAGGACTCGGGGGCGGCCGGCCTTGTGCACTGGCAGGGCCGG

29 FPVTLGS* (ORF29)

GGCGGGTGCGCGCCCTCTCAGACGTACAGGGTGTTGGGCCCCTGACCACACAGCACCCGG

48520 + + + + + + 48579

CCGCCCACGCGCGGGAGAGTCTGCATGTCCCACAACCCGGGGACTGGTGTGTCGTGGGCC

CCGTACAGCTCCAGGTTGGTGCTCGGGTTCATGCAGGTGCAGCGTGATGCTCTGGGCATC

48580 + + + + + + 48639

GGCATGTCGAGGTCCAACCACGAGCCCAAGTACGTCCACGTCGCACTACGAGACCCGTAG
30 (ORF30)* APAAHHEPC

GCTGCACGCGCTGGATCGGGACGTCGTTGTAGATCGAGGACCCGCCGCTCGCCTGGGCGA

48640 + + + + + + 48699

CGACGTGCGCGACCTAGCCCTGCAGCAACATCTAGCTCCTGGGCGGCGAGCGGACCCGCT

30 RQVRQI PVDNYI S SGGSAQA

GGATGTCCACCGACTCCTTGCCCAGTCGGCACGCCCGCCCCAGCAGGCCGCGGCACAGCA

48700 + + + + + + 48759

CCTACAGGTGGCTGAGGAACGGGTCAGCCGTGCGGGCGGGGTCGTCCGGCGCCGTGTCGT

61

30 LIDVSEKGLRCARGLLGRCL

CCCGCTCCTCCAGCGTCCAGGCCTCGCCCGAAGCCCCCTTGGAGTCGACGAGGTCGGCCA

48760 + + + + + + 48819

GGGCGAGGAGGTCGCAGGTCCGGAGCGGGCTTCGGGGGAACCTCAGCTGCTCCAGCCGGT
30 VREELTWAEGSAGKSDVLDA

GCCGATGGGCGTGGAACCGTGCCTCGTCGGCCAGCAGGGTCGCCTCGCCGAGCTGCAGGT

48820 + + + + + + 48879

CGGCTACCCGCACCTTGGCACGGAGCAGCCGGTCGTCCCAGCGGAGCGGCTCGACGTCCA
30 LRHAHFRAEDALLTAEGLQL

GGGTGATCGGCGCCGAGCCCTGCTCCTCGTACTCGGTGTAGGTGATCTTGCGGCCGGGCA

48880 + + + + + + 48939

CCCACTAGCCGCGGCTCGGGACGAGGAGCATGAGCCACATCCACTAGAACGCCGGCCCGT
30 HTI PASGQEEYETYTIKRGP

GCCTCCCGCGGAAGACGTCCTGAGCGGCCGCGGCCAGTCCGGTCATGGTGCCGACCGACG

48940 + + + + + + 48999

CGGAGGGCGCCTTCTGCAGGACTCGCCGGCGCCGGTCAGGCCAGTACCACGGCTGGCTGC
30 LRGRFVDQAAAALGTMTGVS

AGGCCGAGGCCACGGCCAGCATCGGCGCCCGGAACATCGGTGATCCGGCGTTGAGTTCGG

49000 + + + + + + 49059

TCCGGCTCCGGTGCCGGTCGTAGCCGCGGGCCTTGTAGCCACTAGGCCGCAACTCAAGCC
30 SASAVALMPARFMPSGANLE

AGGCGTACTGCTGCTGGAGCACCGCGCCCAGCGGAAGGACGCGCTCCTGGGGAACGAAGA

49060 + + + + + + 49119

TCCGCATGACGACGACCTCGTGGCGCGGGTCGCCTTCCTGCGCGAGGACCCCTTGCTTCT
30 SAYQQQLVAGLPLVREQPVF

CGTCCGCGGCGATGGTGCTGACGCTTCCCGAGCCCCGGAGCCCCGAGGTGTGCCAGTCGT

49120 + + + + + + 49179

GCAGGCGCCGCTACCACGACTGCGAAGGGCTCGGGGCCTCGGGGCTCCACACGGTCAGCA
30 VDAAITSVSGSGRLGSTHWD

CGACGATCTGCAGCTGGTCGGTCGGCACCAGGGCCATCACGGGCTGCATGCCGCCGTCGG

49180 + + + + + + 49239

GCTGCTAGACGTCGACCAGCCAGCCGTGGTCCCGGTAGTGCCCGACGTACGGCGGCAGCC
30 DVIQLQDTPVLAMVPQMGGD

GGGTCGGTGAGACGGCGATCAGAACCTGCCAGTGACTGTGCCAGGCACCGCTGATGAAGC

49240 + + + + + + 49299

CCCAGCCACTCTGCCGCTAGTCTTGGACGGTCACTGACACGGTCCGTGGCGACTACTTCG
30 PTPSVAILVQWHSHWAGSIF

CCCACTTGCCGTTCACTACGACACCGCCGTCGACCGGGGCCGCCATGCCGCCGGGACTGA

49300 + + + + + + 49359

GGGTGAACGGCAAGTGATGCTGTGGCGGCAGCTGGCCCCGGCGGTACGGCGGCCCTGACT
30 GWKGNVVVGGDVPAAMGGPS

GGGTGCCGGAGACCCGGACATCCGGCCGGGAGAACACCTCGTCCTGCACGTGGTCGGGGA

49360 + + + + + + 49419

CCCACGGCCTCTGGGCCTGTAGGCCGGCCCTCTTGTGGAGCAGGACGTGCACCAGCCCCT
30 LTGSVRVDPRSFVEDQVHDP

AGAGGCCCGCCATCCAGGTGGGTATCCACCACACCGAGGCCGTCCAGGCGGCCGATCCGT

49420 + + + + + + 49479

TCTCCGGGCGGTAGGTCCACCCATAGGTGGTGTGGCTCCGGCAGGTCCGCCGGCTAGGCA
30 FLGAMWTP IWWVSATWAASG

CGCCGCGCGCCAGCTCGGCGGCCACGTCCACCAGGGTGCGGGCGTCGGACTCGAAGCCGC

49480 + + + + + + 49539

GCGGCGCGCGGTCGAGCCGCCGGTGCAGGTGGTCCCACGCCCGCAGCCTGAGCTTCGGCG
30 DGRALEAAVDVLTRADSEFG

62

CGTAACGGGCCGGCACGCGCATGCGGAAGATCCCGGCTTCGGCCATCGCCTCGACCGACT

49540 + + + + + + 49599

GCATTGCCCGGCCGTGCGCGTACGCCTTCTAGGGCCGAAGCCGGTAGCGGAGCTGGCTGA
30 GYRAPVRMRFI GAEAMAEVS

CCTCGTGCAGCCGCCGGTTCTCCTCGGTCCAGGCCGCGTGGGACTGGAGCAGCGGCCTCA

49600 + + + + + + 49659

GGAGCACGTCGGCGGCCAAGAGGAGCCAGGTCCGGCGCACCCTGACCTCGTCGCCGGAGT
30 EEHLRRNEETWAAHSQLLPR

GCTTCGAGGCCCGTTCCACCAGTTCGGTACGGGCGGGCGTAGACGTCTGGTCCACTCGAT

49660 + + + + + + 49719

CGAAGCTCCGGGCAAGGTGGTCAAGCCATGCCCGCCCGCATCTGCAGACCAGGTGAGCTA

30 LKSAREVLETRAPTSTQDV
(ORF3 0)

CCTCCAGGAATCATGAGACGCCCTGTCCGCGGTATGCGGAAGCAGGCGTCTGCGCGCATC

49720 + + + + + + 49779

GGAGGTCCTTAGTACTCTGCGGGACAGGCGCCATACGCCTTCGTCCGCAGACGCGCGTAG

GGTCAGGACGGCGTCGCCCTGCTCCCGCATGGTTCACCGAGTTCCGCGGACGTCGCATCT

49780 + + + + + + 49839

CCAGTCCTGCCGCAGCGGGACGAGGGCGTACCAAGTGGCTCAAGGCGCCTGCAGCGTAGA

CCTTGATTGCCGGTCACCTACCCCGATGCCGATCGGGCTGGTGCGACAGCGCATCCCACG

49840 + + + + + + 49899

GGAACTAACGGCCAGTGGATGGGGCTACGGCTAGCCCGACCACGCTGTCGCGTAGGGTGC

AGAAGTCCACGAACGGTCCGGGAAGCCAGAATGTGCTTCTCGGCCGGAGTCACGGCCGGC

49900 + + + + + + 49959

TCTTCAGGTGCTTGCCAGGCCCTTCGGTCTTACACGAAGAGCCGGCCTCAGTGCCGGCCG

GCCGGCGCCCGTCGCCGGTCACGCCGGACCACGCCCGGACCGGTCATGGAGGCAGCCCAT

49960 + + + + + + 50019

CGGCCGCGGGCAGCGGCCAGTGCGGCCTGGTGCGGGCCTGGCCAGTACCTCCGTCGGGTA

GAGTGACAACGACAGTCCGTCCCGGGTGCCGGCCGCGGTGGCACCCGCCACCGCGAAACC

50020 + + + + + + 50079

CTCACTGTTGCTGTCAGGCAGGGCCCACGGCCGGCGCCACCGTGGGCGGTGGCGCTTTGG

GTCGGCCGGCACGGTCCTCGGCGCCGCGGTGGCTTCGCCCGCCGCCTACACCGCGGCGAC

50080 + + + + + + 50139

CAGCCGGCCGTGCCAGGAGCCGCGGCGCCACCGAAGCGGGCGGCGGATGTGGCGCCGCTG

CGCCCAGGAAGCGGCGACCGCGCTGGTCCGCATGCTGATGGAACAGATGGTGCTCGGTCC

50140 + + + + + + 50199

GCGGGTCCTTCGCCGCTGGCGCGACCAGGCGTACGACTACCTTGTCTACCACGAGCCAGG

CGGCGCGGTCGGTCCCGAGACCCGCGCGGACGGCCCGGCGGGGCGGACCGGCTCCGGCCA

50200 + + + + + + 50259

GCCGCGCCAGCCAGGGCTCTGGGCGCGCCTGCCGGGCCGCGCCGCCTGGCCGAGGCCGGT

CGGCCCGGCGCCGCAGACCGGACCGGACGCGCCGGGCGAACCCCCGCCCACGTGGGCGCC

50260 + + + + + + 50319

GCCGGGCCGCGGCGTCTGGCCTGGCCTGCGCGGCCCGCTTGGGGGCGGGTGCACCCGCGG

GAACCTCGACGACGGGAAGGTAGGAGGACGATGAGGCCGCTCGTTCGGGCAGTGCTGCGG

50320 + + + + + + 50379

CTTGGAGCTGCTGCCCTTCCATCCTCCTGCTACTCCGGCGAGCAAGCCCGTCACGACGCC
31 (ORF31) MRPLVRAVLR-

GGTTCCCTGCGGCAGGTGAGGTACGTGGACGTGGTCTCCCCGCGCCGGGCGCGCTCCCTG

50380 + + + + + + 50439

CCAAGGGACGCCGTCCACTCCATGCACCTGCACCAGAGGGGCGCGGCCCGCGCGAGGGAC

31 GSLRQVRYVDVVSPRRARSL

63

GTGGCGCGGGTGTACCGGGAGACCGAGGAGCAGTTCGGCGTGCTCGCGCCCCCCCTGGCC

50440 + + + + + + 50499

CACCGCGCCCACATGGCCCTCTGGCTCCTCGTCAAGCCGCACGAGCGCGGGGGGGACCGG
31 VARVYRETEEQFGVLAPPLA

CTCCACTCGCCCGCCGCGGCGTCGCTGGCCGCGACGTGGCTCATGCTGCGGGAGACACTG

50500 + + + + + + 50559

GAGGTGAGCGGGCGGCGCCGCAGCGACCGGCGCTGCACCGAGTACGACGCCCTCTGTGAC
31 LHSPAAASLAATWLMLRETL

CTGGTCGACGGGCGGGTGAGCCGGGCGGTGAAGGAGACGGTCGCCACCGAGGTCTCCCGT

50560 + + + + + + 50619

GACCAGCTGCCCGCCCACTCGGCCCGCCACTTCCTCTGCCAGCGGTGGCTCCAGAGGGCA
31 LVDGRVSRAVKETVATEVSR

GCCAACGACTGTCCGTACTGCGTCCAGGTCCATCAGGCGGTACTCGGGACACTGCCTCCG

50620 + + + + + + 50679

CGGTTGCTGACAGGCATGACGCAGGTCCAGGTAGTCCGCCATGAGCCCTGTGACGGAGGC
31 ANDCPYCVQVHQAVLGTLPP

GACGGCGGCCAGGCCGGGCTCCTGCGGTGGGTCCGGGAGGCAGGCCGACGGCCCGGCGGC

50680 + + + + + + 50739

CTGCCGCCGGTCCGGCCCGAGGACGCCACCCAGGCCCTCCGTCCGGCTGCCGGGCCGCCG
31 DGGQAGLLRWVREAGRRPGG

GGTGCGGTGGGCGGCGGGCGGCCGCTTCCGTTCAGCGGTGAACAGGCACCGGAACTGTGC

50740 + + + + + + 50799

CCACGCCACCCGCCGCCCGCCGGCGAAGGCAAGTCGCCACTTGTCCGTGGCCTTGACACG
31 GAVGGGRPLPFSGEQAPELC

GGCGTCGTGGTCACGTTCCACTACATCAACCGCATGGTCTCCCTCTTCCTCGACGACTCC

50800 + + + + + + 50859

CCGCAGCACCAGTGCAAGGTGATGTAGTTGGCGTACCAGAGGGAGAAGGAGCTGCTGAGG
31 GVVVTFHYINRMVSLFLDDS

CCCATGCCGACCCGGACGCCGACACCGTTGCGCGGGCCCATCATGAGGACCACCGCACTG

50860 + + + + + + 50919

GGGTACGGCTGGGCCTGCGGCTGTGGCAACGCGCCCGGGTAGTACTCCTGGTGGCGTGAC
31 PMPTRTPTPLRGPIMRTTAL

GCCATGCGTCCCGTCGGCCCGGGGCTGCTGACACCGGGCGCATCGCTCGGCCTGCTGCCT

50920 + + + + + + 50979

CGGTACGCAGGGCAGCCGGGCCCCGACGACTGTGGCCCGCGTAGCGAGCCGGACGACGGA
31 AMRPVGPGLLTPGASLGLLP

CCGGCTCCCCTGCCGCCCGGACTGGAGTGGGCCGAGGGCAACCCTTTCGTGGCCCAGGCG

50980 + + + + + + 51039

GGCCGAGGGGACGGCGGGCCTGACCTCACCCGGCTCCCGTTGGGAAAGCACCGGGTCCGC
31 PAPLPPGLEWAEGNPFVAQA

CTGGGGCGTGCCGTCGCCGCTGTGGACCAGGGAGCGCACTGGGTGCCCGAACCGGTCCGG

51040 + + + + + + 51099

GACCCCGCACGGCAGCGGCGACACCTGGTCCCTCGCGTGACCCACGGGCTTGGCCAGGCC
31 LGRAVAAVDQGAHWVPEPVR

GAGCGGCTGCGCACACGTCTGGACACCTGGGACGGATCGGCGCCGGGCCTCGGCCGGGGA

51100 + + + + + + 51159

CTCGCCGACGCGTGTGCAGACCTGTGGACCCTGCCTAGCCGCGGCCCGGAGCCGGCCCCT
31 ERLRTRLDTWDGSAPGLGRG

TGGCTCGACGAGGCCGTGTCCGGCCTGCCGCCCCAGGACGTGCCCGCGGCACGGCTGGCG

51160 + + + + + + 51219

ACCGAGCTGCTCCGGCACAGGCCGGACGGCGGGGTCCTGCACGGGCGCCGTGCCGACCGC
31 WLDEAVSGLPPQDVPAARLA

CTGCTGACGGCCTTCGCCCCCTACCAGGTGCTCCCGGACGACGTCGAGGAGTTCAGACGG

64

51220 + + + + + + 51279

GACGACTGCCGGAAGCGGGGGATGGTCCACGAGGGCCTGCTGCAGCTCCTCAAGTCTGCC
31 LLTAFAPYQVLPDDVEEFRR

CGTCGGCCCACCGACCGCGAACTCGTCGAGCTCACGTCCTACGCCGCGCTGACCACGGCC

51280 + + + + + + 51339

GCAGCCGGGTGGCTGGCGCTTGAGCAGCTCGAGTGCAGGATGCGGCGCGACTGGTGCCGG
31 RRPTDRELVELTSYAALTTA

GTCCGTGTCGGTCGCACGCTCGTCGTGCCCGACGCCGCCGGGCCGGGATGAACGGCCCCG

51340 + + + + + + 51399

CAGGCACAGCCAGCGTGCGAGCAGCACGGGCTGCGGCGGCCCGGCCCTACTTGCCGGGGC

31 VRVGRTLVVPDAAGPG* (ORF31)

CAACGGCTCGGGAAGGCTGTCTCACGGCCGGAGGCGTACGCCGGTGAGGTGCTCGGACTC

51400 + + + + + + 51459

GTTGCCGAGCCCTTCCGACAGAGTGCCGGCCTCCGCATGCGGCCACTCCACGAGCCTGAG

(ORF32) * PRLRVGTLHE S E -

CTCCCAGAGGCGGCGCCGGGCCCTGGGGTCGACGGCTGCTCCGCCGGGGCGCACGAGCCC

51460 + + + + + + 51519

GAGGGTCTCCGCCGCGGCCCGGGACCCCAGCTGCCGACGAGGCGGCCCCGCGTGCTCGGG

32 EWLRRRARPDVAAGGPRVLG-

GGGTGCGCCCCGGGTCTCGGTCACGCCGAGGGGCCCGTAGAACTCGCCCCCGCGCGCGCC

51520 + + + + + + 51579

CCCACGCGGGGCCCAGAGCCAGTGCGGCTCCCCGGGCATCTTGAGCGGGGGCGCGCGCGG
32 PAGRTETVGLPGYFEGGRAG-

GGGATCGGTGGCCGCCCGCAGACCAGGCAGCATCCCCGCCGCGGCGGGCTGCAGGAACAA

51580 + + + + + + 51639

CCCTAGCCACCGGCGGGCGTCTGGTCCGTCGTAGGGGCGGCGCCGCCCGACGTCCTTGTT
32 PDTAARLGPLMGAAAPQLFL-

CGGGGCGAGCGGGGAGCCGAGCCTGCGCACGGGCGCGGGAAAGTCCCGGCCCAGACCGGT

51640 + + + + + + 51699

GCCCCGCTCGCCCCTCGGCTCGGACGCGTGCCCGCGCCCTTTCAGGGCCGGGTCTGGCCA
32 PAL PSGLRRVPAPFDRGLGT-

CGCGGTCAGCCCGGGATGAGCGGCGAGCGAGGCCAGTTCCGCGCCGGACTCCGCCAGTCT

51700 + + + + + + 51759

GCGCCAGTCGGGCCCTACTCGCCGCTCGCTCCGGTCAAGGCGCGGCCTGAGGCGGTCAGA
32 ATLGPHAALSALEAGS EALR-

GTGATGGAGTTCCAGCGCGAACATGAGGTTGGCCAGCTTGGACTGGTTGTAGGCCCGGTA

51760 + + + + + + 51819

CACTACCTCAAGGTCGCGCTTGTACTCCAACCGGTCGAACCTGACCAACATCCGGGCCAT
32 HHLELAFMLNALKS QNYARY-

CCGGCTGTAGCGGCGTTCGCCGTGAAGGTCGCTGAAGTCGATGCGCCCCAGCCGGTGCAG

51820 + + + + + + 51879

GGCCGACATCGCCGCAAGCGGCACTTCCAGCGACTTCAGCTACGCGGGGTCGGCCACGTC
32 RSYRREGHLDS FD I RGLRHL-

ATAGCTGCTGATCGTCACGACCCGCGCGCCCGGCGCGGCCCGCAGGCTGTCCAGGAGCAG

51880 + + + + + + 51939

TATCGACGACTAGCAGTGCTGGGCGCGCGGGCCGCGCCGGGCGTCCGACAGGTCCTCGTC
32 YSSITVVRAGPAARLSDLLL-

GCCGGTGAGGGCGAAGTGCCCCAGGTGGTTCGTGGCGAACTGGAGTTCGTGACCGTCCGG

51940 + + + + + + 51999

CGGCCACTCCCGCTTCACGGGGTCCACCAAGCACCGCTTGACCTCAAGCACTGGCAGGCC
32 GTLAFHGLHNTAFQLEHGDP-

GGTGCGGGCCCGGTCGGTCCACATCACGCCCGCGTTGTTGACCAGCAGGTGGATGCGCGG
52000 + + + + + + 52059

65

CCACGCCCGGGCCAGCCAGGTGTAGTGCGGGCGCAACAACTGGTCGTCCACCTACGCGCC

TRARDTWMVGANNVLLH I R P -

GAAGCGGTCGCGCAGTTCCTCGGCGCCGGCACGCACCGACGCGAGACGGGAAAGATCCAG

52060 + + + + + + 52119

CTTCGCCAGCGCGTCAAGGAGCCGCGGCCGTGCGTGGCTGCGCTCTGCCCTTTCTAGGTC

FRDRLEEAGARVSALRSLDL-

CCGTCTGACCGTCAGTTGCGCCGACGGCACCCGGCTTTGGATGCGGGCCGCCGCGGCGAC

52120 + + + + + + 52179

GGCAGACTGGCAGTCAACGCGGCTGCCGTGGGCCGAAACCTACGCCCGGCGGCGCCGCTG

RRVTLQAS PVRS Q I R A A A A V -

CCCGCGGTCCGGATCGCGCACGGCCAGCACCACGTGGGCGCCGTGCCGGGCGAGCTCCTG

52180 + + + + + + 52239

GGGCGCCAGGCCTAGCGCGTGCCGGTCGTGGTGCACCCGCGGCACGGCCCGCTCGAGGAC

GRD PDRVALVVHAGHRALE Q -

CGCCAGGTGCAGTCCGATGCCGGAGCTGGCACCGGTGACCACCGCGGTGGTTCCGGTACG

52240 + + + + + + 52299

GCGGTCCACGTCAGGCTACGGCCTCGACCGTGGCCACTGGTGGCGCCACCAAGGCCATGC

ALHLG I GS SAGTVVATTGTR-

GTCCGGGACATCGGCGGCGCTCCAGCGTCGCCGCGTTCTCATCGGTCGTCCCTCCCGGGG

52300 4- + + + + + 52359

CAGGCCCTGTAGCCGCCGCGAGGTCGCAGCGGCGCAAGAGTAGCCAGCAGGGAGGGCCCC
DPVDAASWRRRTRM (ORF32)

GATGCGTCAGCCGGCCTGGGCCATCGCGGCCCGGTAGCCGTTGGCGACGATCTGCCGGGC

+ + + + + h

CTACGCAGTCGGCCGGACCCGGTAGCGCCGGGCCATCGGCAACCGCTGCTAGACGGCCCG

GGAGTGCTCGTAGTACTCGTCGTCCTTCGGCAGCTCCGTGGCGAGACCGCTGACGTACCG

52420 + + + + + + 52479

CCTCACGAGCATCATGAGCAGCAGGAAGCCGTCGAGGCACCGCTCTGGCGACTGCATGGC

GTTGAACATGCAGAACGCGGCGGCGATCAGAACGGTGTCGTGCAGAGCGGTGTCGTCCGC

52480 + + + + + + 52539

CAACTTGTACGTCTTGCGCCGCCGCTAGTCTTGCCACAGCACGTCTCGCCACAGCAGGCG

TCCCTCGGCCCGCGCCGAGGCGATCACCCCTGCGGAGACCGGGCGCGCCGCGCTCTGGAC

52540 + + + + + + 52599

AGGGAGCCGGGCGCGGCTCCGCTAGTGGGGACGCCTCTGGCCCGCGCGGCGCGAGACCTG

CTCGGCGGCGACGGCCAGCAGCGCGCGCGTCCTGCCGTCGATGGGCGCGGTGGCGGGGTC

GAGCCGCCGCTGCCGGTCGTCGCGCGCGCAGGACGGCAGCTACCCGCGCCACCGCCCCAG

GGCGAGGACGGCCTCGACGAGCTGCCGGCCTCCCGGCAGCTGCGCGGCGGCGAAGGCCCC

+ 1. h + + +

CCGCTCCTGCCGGAGCTGCTCGACGGCCGGAGGGCCGTCGACGCGCCGCCGCTTCCGGGG

GTGGGAGGCGGCGCAGAACTCGGTGGAGTTGAGATGCGAGACGTACGCCGCGATGAGCTC

52720 + + + + + + 52779

CACCCTCCGCCGCGTCTTGAGCCACCTCAACTCTACGCTCTGCATGCGGCGCTACTCGAG

GCGTTGCCCCGGTTCCAGCGAGGACGGCGCCCGCAGCAGGGCGTTCGCGAGATCGCCCAG

52780 + + + + + + 52839

CGCAACGGGGCCAAGGTCGCTCCTGCCGCGGGCGTCGTCCCGCAAGCGCTCTAGCGGGTC

CGGTGCTGCGGTGCCGGGGTGGTGAGCCATCAGACCACTGATGCCGGGGAGGTCGTTGTC

52840 + + + + + + 52899

GCCACGACGCCACGGCCCCACCACTCGGTAGTCTGGTGACTACGGCCCCTCCAGCAACAG

GAGTGCTATGTGGGGCACGGCTCTTCCTTCCGGGTGGACGAGGGGCGGACGGCGGCGGAT
52900 + + + + + + 52959

66

CTCACGATACACCCCGTGCCGAGAAGGAAGGCCCACCTGCTCCCCGCCTGCCGCCGCCTA

CAGGGCCATTCGACTTCGTCGTCGGCGGCCGCGCAGATGCGGGTGAAGGGCCATTCCACG

5 29 6 o + + + + + + 53019

GTCCCGGTAAGCTGAAGCAGCAGCCGCCGGCGCGTCTACGCCCACTTCCCGGTAAGGTGC

TCTTCCCCTCCCGTTGCGGAGTGGGCGGAGGCCGTGGTGAAGAGGGTGACGAGTCCGAAC

53020 + + + + + + 53079

AGAAGGGGAGGGCAACGCCTCACCCGCCTCCGGCACCACTTCTCCCACTGCTCAGGCTTG

GTGCCGAAGAGGAGGGACAGTCGGGCAACGTGAAGTGCGGTACCCATGCGAGCTCCTAGC

53080 + + + + + + 53139

CACGGCTTCTCCTCCCTGTCAGCCCGTTGCACTTCACGCCATGGGTACGCTCGAGGATCG

GAGGGCGGCGTGACCGCGGGACGGTGAGACCTCGTGATGCCAGGAAGCTAGCGAATCGGA

53140 + + + + + + 53 1"

CTCCCGCCGCACTGGCGCCCTGCCACTCTGGAGCACTACGGTCCTTCGATCGCTTAGCCT

CTGAGGGTGGCAACGATATGCCAGACTTTGGCAACTTGCCTGTGTATCAGCCGGACTGTC

53200 + + + + + + 53259

GACTCCCACCGTTGCTATACGGTCTGAAACCGTTGAACGGACACATAGTCGGCCTGACAG

(ORF33) V Y Q P D C R

GGCCGCTGGTAAAGACGGAACGGCGAGATCCCGCGACCGCGTCGCAGAGCAGCAGGGTCT

53260 + + + + + + 53319

CCGGCGACCATTTCTGCCTTGCCGCTCTAGGGCGCTGGCGCAGCGTCTCGTCGTCCCAGA

PLVKTERRDPATASQSSRVC-

GCTCACCCAGCGTCGGGGCGGCCAGCATGTCGCGTACCGGGAGCGTGACGCCCAGCTCGC

53320 + + + + + + 53379

CGAGTGGGTCGCAGCCCCGCCGGTCGTACAGCGCATGGCCCTCGCACTGCGGGTCGAGCG

SPSVGAASMSRTGSVTPSSR-

GGTTGATCCTGCGGACCAGCCGGGTGATGAGCAGGGAGTCGCCGCCGTGGGCGAAGAAAT

53380 + + + + + + 53439

■CCAACTAGGACGCCTGGTCGGCCCACTACTCGTCCCTCAGCGGCGGCACCCGCTTCTTTA

LI LRTSRVMS RES P PWAKKS-

CAGCACCTTCGGAGGGGTCCGGGAAGCCGAGCAGGTCACCCCAGCCGCGCACCAGTACCT

53440 + + + + + + 53499

GTCGTGGAAGCCTCCCCAGGCCCTTCGGCTCGTCCAGTGGGGTCGGCGCGTGGTCATGGA

APSEGSGKPSRSPQPRTSTW-

GGCGGATGTCGCCGGTGGTGACGACCGTGCGCCGGGAGCCCCGACGTGCCGAGCGCAGCC

53500 + + + + + + ^-5^=>^

CCGCCTACAGCGGCCACCACTGCTGGCACGCGGCCCTCGGGGCTGCACGGCTCGCGTCGG

RMS PVVTTVRRE PRRAERS R -

GCGAGGCATGCACCAGCGCCACCTGGTCGCCGAGGTTGCGCCGCGACAGCTCGCGCAGCG

53560 + + + + + + 53619

CGCTCCGTACGTGGTCGCGGTGGACCAGCGGCTCCAACGCGGCGCTGTCGAGCGCGTCGC

EACTSATWS PRLRRD S S RSD-

ACACCGTGACGCCGAACCTCTCGGTGATCCTGCGGACCAGCCGCGTGATCAGCAGCGTGT

53620 + + + + + + 53679

TGTGGCACTGCGGCTTGGAGAGCCACTAGGACGCCTGGTCGGCGCACTAGTCGTCGCACA

TVTPNLSVI LRTSRVI S SVS-

CCCCGCCGCGCGCGAAGAAATCCGAATGCTCGGTGAGGTCGGAGCGGCCGAGGAGCTCGC

53680 + + + + + + 53739

GGGGCGGCGCGCGCTTCTTTAGGCTTACGAGCCACTCCAGCCTCGCCGGCTCCTCGAGCG

PPRAKKSECSVRSERPRSSL-

TCCACGCGCCGACCATGAACTCCCCCACGTCACCGAGCCGGTGCTCGTCGCCGTCGGGGC

53740 + + + + + + 53799

AGGTGCGCGGCTGGTACTTGAGGGGGTGCAGTGGCTCGGCCACGAGCAGCGGCAGCCCCG

67

HAP

TMNSPTSPSRCSSPSGP-

CCTTCGGCGCGCCGGATCCCGCGGAACGGTTCCGGCCGGAGACGGCAGAGCGGTCACTGG

53800 + + + + + + 53859

GGAAGCCGCGCGGCCTAGGGCGCCTTGCCAAGGCCGGCCTCTGCCGTCTCGCCAGTGACC
FGAPDPAERFRPETAERSLV-

TCACTTTCGCCACCTCCAGGGGCATGTGTCGGCTGCATCGGCTTCCCGCCACGGTACGGG

53860 + + + + + + 53919

AGTGAAAGCGGTGGAGGTCCCCGTACACAGCCGACGTAGCCGAAGGGCGGTGCCATGCCC

TFATSRGMCRLHRL PATVRE-

AGCACATGTTGCATGGCAATACCTTTCCAAGTCGGTGGCAACCCTCCTTGCCATCCACCC

53920 + + + + + + 53979

TCGTGTACAACGTACCGTTATGGAAAGGTTCAGCCACCGTTGGGAGGAACGGTAGGTGGG

HMLHGNTFPSRWQPSLPSTH-

ACTGCAGTTGGGCGAGATGTGTAGGCATTCGAGGTCCGCAGGTTTGCCAAGCCGCGCGCG

53980 + + + + + + 54039

TGACGTCAACCCGCTCTACACATCCGTAAGCTCCAGGCGTCCAAACGGTTCGGCGCGCGC

C SWARCVGI RGPQVCQAARD-

ACCGGCATACTCTCTGGCACAACTGGAATGAGTAGCGTGGCAGGCCACGGGGACCGGGCC

54040 + + + + + + 54099

TGGCCGTATGAGAGACCGTGTTGACCTTACTCATCGCACCGTCCGGTGCCCCTGGCCCGG

RHTLWHNWNE* ( ORF3 3 )

GGGCCAGGAACCTTCGTCCTCCATCTATTCGCTGGGGCGTGCACGTGTTGGAGCAGCCAT

54100 + + + + + + 54159

CCCGGTCCTTGGAAGCAGGAGGTAGATAAGCGACCCCGCACGTGCACAACCTCGTCGGTA

CTTTCGGCCGTCGCCTGAGGCAGCTGAGGACCGAGCGGGGTCTTTCCCAGGCCGCGCTCG

54160 + + + + + + 54219

GAAAGCCGGCAGCGGACTCCGTCGACTCCTGGCTCGCCCCAGAAAGGGTCCGGCGCGAGC

CGGGGGACGGCATGTCTACGGGCTATCTCTCGCGCCTGGAGTCGGGCGCCCGGCAGCCCT

54220 + + + + + + 54279

GCCCCCTGCCGTACAGATGCCCGATAGAGAGCGCGGACCTCAGCCCGCGGGCCGTCGGGA

(ORF34) MSTGYLSRLESGARQPS-

CCGATCGCGCCGTCGCCCACCTGGCCGGACAACTCGGCATCAGCCCGTCGGAGTTCGAAG

54280 + + + + + + 54339

GGCTAGCGCGGCAGCGGGTGGACCGGCCTGTTGAGCCGTAGTCGGGCAGCCTCAAGCTTC

DRAVAHLAGQLGI SPSEFEG-

GGTCCCGGGCCACCTCGCTCGCCCAGATCCTCTCCCTCTCCACTTCCCTGGAGTCCGACG

54340 + + + + + + 54399

CCAGGGCCCGGTGGAGCGAGCGGGTCTAGGAGAGGGAGAGGTGAAGGGACCTCAGGCTGC

SRATSLAQILSLSTSLESDE-

AGACCAGTGAGCTTCTCGCCGAGGCGGTACGTTCCGCGCATGGCCAGGATCCGATGCTCC

54400 + + + + + + 54459

TCTGGTCACTCGAAGAGCGGCTCCGCCATGCAAGGCGCGTACCGGTCCTAGGCTACGAGG

TS ELLAEAVRSAHGQ D PMLR-

GCTGGCAGGCCCTGTGGCTGCTGGGACAGTGGAAGCGCCGGCACGGCGACTCGGCCGGCG

54460 + + + + + + 54519

CGACCGTCCGGGACACCGACGACCCTGTCACCTTCGCGGCCGTGCCGCTGAGCCGGCCGC

WQALWLLGQWKRRHGD SAGE-

AGCACGGCTACCTCCAGCGTCTGGTGACGCTGAGTGAGGAGATCGGCCTGGCCGAGTTGC

54520 + + + + + + 54579

TCGTGCCGATGGAGGTCGCAGACCACTGCGACTCACTCCTCTAGCCGGACCGGCTCAACG

HGYLQRLVTLSEE I GLAELR-

GCGCACGGGCCCTGACCCAGTTCGCCCGGTCGCTGCGGGTACTGGGCGAGATCGTTCCGG

68

54580 + + + + + + 54639

CGCGTGCCCGGGACTGGGTCAAGCGGGCCAGCGACGCCCATGACCCGCTCTAGCAAGGCC
34 ARALTQFARS LRVLGE I V P A -

CGGTGGAGGCTGCCGCCGCCGCCCACCGGCTCGCGGTGGACCATGCGCTGTCCAGCCAGG

54640 + + + + + + 54699

GCCACCTCCGACGGCGGCGGCGGGTGGCCGAGCGCCACCTGGTACGCGACAGGTCGGTCC
34 VEAAAAAHRLAVDHALSSQD-

ACAGGGCCGCTTCGCTGCTGGTTCTGGTGTCGGTGGAGGCCGAGGCGGGACGGATGCCCG

54700 + + + + + + 54759

TGTCCCGGCGAAGCGACGACCAAGACCACAGCCACCTCCGGCTCCGCCCTGCCTACGGGC
34 RAASLLVLVSVEAEAGRMPD-

ACGCCCGGCGCCACGCCGACGAACTGACCGTCCTGGTGAGGGGACGGTCCGACACTCTGT

54760 + + + + + + 54819

TGCGGGCCGCGGTGCGGCTGCTTGACTGGCAGGACCACTCCCCTGCCAGGCTGTGAGACA
34 ARRHADELTVLVRGRSDTLW-

GGGCCGAGGCGTTGTGGACGGCGGGTGCGTTGAAGGTGCGGCAGGGCGAGTTCGCCGCGG

54820 + + + + + + 54879

CCCGGCTCCGCAACACCTGCCGCCCACGCAACTTCCACGCCGTCCCGCTCAAGCGGCGCC
34 AEALWTAGALKVRQGE F A A A -

CCGAGGTCCTTTTCCAGGAGGCTCTGGACGGGTTCGACAGCCGGGAGAACCTGACGATCT

54880 + + + + + + 54939

GGCTCCAGGAAAAGGTCCTCCGAGACCTGCCCAAGCTGTCGGCCCTCTTGGACTGCTAGA
34 EVLFQEALDGFDSRENLTIW-

GGCTGCGGCTGCGCATCGCGATGGCCGAACTCCACCTGCAGAAACTTCCTCCCGAGCCCG

54940 + + + + + + 54999

CCGACGCCGACGCGTAGCGCTACCGGCTTGAGGTGGACGTCTTTGAAGGAGGGCTCGGGC
34 LRLRIAMAELHLQKLPPEPD-

ACGCCGCGCAGCTCTGCATCGAGGCGGCGGAGGCGGCCCTTCCCTTTGCCCGCACATCCG

55000 + + + + + + 55059

TGCGGCGCGTCGAGACGTAGCTCCGCCGCCTCCGCCGGGAAGGGAAACGGGCGTGTAGGC
34 AAQLCIEAAEAALPFARTSA-

CTCTGGAACAGTCCCTCGCCGCTCTGCGGGCGCGCCTCGCCTTCCATGAGGGCAGGTTCG

55060 + + + + + + 55119

GAGACCTTGTCAGGGAGCGGCGAGACGCCCGCGCGGAGCGGAAGGTACTCCCGTCCAAGC
34 LEQS LAALRARLAFHEGRFA-

CCGATGCCCGCGCGTTGTTGGAGAGGCTCGGCAGGACCGAGCTCCGGCTGCCCTATCAGA

55120 + + + + + + 55179

GGCTACGGGCGCGCAACAACCTCTCCGAGCCGTCCTGGCTCGAGGCCGACGGGATAGTCT
34 DARALLERLGRTELRL P Y Q S -

GCCGGATCCGCCTGGAGGTCCTCGGTCATCAGCTGCGCATCCTGAGCGGGGAGGAGGAGG

55180 + + + + + + 55239

CGGCCTAGGCGGACCTCCAGGAGCCAGTAGTCGACGCGTAGGACTCGCCCCTCCTCCTCC
34 RIRLEVLGHQLRILSGEEEE-

AAGGCCTGGCCGGCCTCCAGCTCCTGGCCGAGGAGGCGCAGGAGAACTCCAACATCAACC

55240 + + + + + + 55299

TTCCGGACCGGCCGGAGGTCGAGGACCGGCTCCTCCGCGTCCTCTTGAGGTTGTAGTTGG
34 GLAGLQLLAEEAQENSNINL-

TCGCCGCGGAGATCTGGCGGCTCGCGGCGGAATGCCTGATGCGGGCGCGCGGGAAGGTCC

55300 + + + -f + + 55359

AGCGGCGCCTCTAGACCGCCGAGCGCCGCCTTACGGACTACGCCCGCGCGCCCTTCCAGG
34 AAE1WRLAAECLMRARGKVR-

GCGGCGCCACCGGCGGCTGACGCCGCGCCGGTTCGCGAGGTCCACCGCGCCGCCGTGGCC
55360 + + + + + + 55419

69

CGCCGCGGTGGCCGCCGACTGCGGCGCGGCCAAGCGCTCCAGGTGGCGCGGCGGCACCGG

34 G A T G G * (ORF34)

ACCGCCGTCGGCGTGAGGCGCCGGCGTGTGCCGCCCCCCACGGTTGCTCGCCCTTGGTGG

55420 + + + + + + 55479

TGGCGGCAGCCGCACTCCGCGGCCGCACACGGCGGGGGGTGCCAACGAGCGGGAACCACC

TGCATCTGTTGGCACATGTGTACCTCCTACACAGTCAATTGTTGCCAAAATTGTCGAACC

55480 + -f + + + + 55539

ACGTAGACAACCGTGTACACATGGAGGATGTGTCAGTTAACAACGGTTTTAACAGCTTGG

GAATGGCAATTGCTTGCCTTTGCTGAAGAGGCGTGCTGATATGCAAGTCAAGTAGCCTCC

55540 + + + + + + 55599

CTTACCGTTAACGAACGGAAACGACTTCTCCGCACGACTATACGTTCAGTTCATCGGAGG

TC CGAT CT CGGGCGGC CATATGGGAAACATCGAGTTGAGCGGCGATGGCGTTCGTCAGTG

55600 + + + + 4- + 55659

AGGCTAGAGCCCGCCGGTATACCCTTTGTAGCTCAACTCGCCGCTACCGCAAGCAGTCAC

CTGCCGTTCTGGCCAGGCAACTGATGTCGATGGGGATGGCAAGATTTTGCCGAAAACCGA

55660 + + + + + + 55719

GACGGCAAGACCGGTCCGTTGACTACAGCTACCCCTACCGTTCTAAAACGGCTTTTGGCT

TACATCTCTGTCCGTCCCGGACAGCCTTCGCCCCCCGGGTGACACTGCTCCGGCATGGCT

55720 + + + + + + 55779

ATGTAGAGACAGGCAGGGCCTGTCGGAAGCGGGGGGCCCACTGTGACGAGGCCGTACCGA

CCGGTTTCTCGTCGCCCGGCCGACGGACCGCACCGTCCGGAACGAGGCGCCGGTGTGCGT

55780 + + + + + + 55839

GGCCAAAGAGCAGCGGGCCGGCTGCCTGGCGTGGCAGGCCTTGCTCCGCGGCCACACGCA

CCGCTGATGGGCACAGCGGCCTCGGCCGCAGCAGGTTCCCACCGAGAAGAATGCCGAGGC

55840 + + + + + + 55899

GGCGACTACCCGTGTCGCCGGAGCCGGCGTCGTCCAAGGGTGGCTCTTCTTACGGCTCCG

CCAGCCGTGAACCACGACATGTCCCAGCGTGCCTTGCTGGAGGCGGCGGCCGAGGGGCTG

55900 + + + + + + 55959

GGTCGGCACTTGGTGCTGTACAGGGTCGCACGGAACGACCTCCGCCGCCGGCTCCCCGAC

CGGCGGCTGGCCGGCGACGCGCGGTGCCGGAGCGCGTCGGCCGCGCCCTCCTCGGCATTG

55960 + + + + + + 56019

GCCGCCGACCGGCCGCTGCGCGCCACGGCCTCGCGCAGCCGGCGCGGGAGGAGCCGTAAC

AGGGACATGTTCTCCCCCGCCGCCCGCCGGTACGTGCTCGCCTCGGACCGCGCGGGGTTC

56020 + + + + + + 56079

TCCCTGTACAAGAGGGGGCGGCGGGCGGCCATGCACGAGCGGAGCCTGGCGCGCCCCAAG

35 CORF35) MFSPAARRYVLASDRAGF

TTCGAGCAGGCTGTCCGGCTGCGCTCCCGGGGGTACCGGGTGAGCGCGGAGTTCGTCGGC

56080 + -f + + + + 56139

AAGCTCGTCCGACAGGCCGACGCGAGGGCCCCCATGGCCCACTCGCGCCTCAAGCAGCCG
35 FEQAVRLRSRGYRVSAEFVG

CCCGATCAGGGAGCCACCGACGCCCTCCACGCGGAGCACGTGGTCGAAGAGCACCTGAGG

56140 + + + + + + 56199

GGGCTAGTCCCTCGGTGGCTGCGGGAGGTGCGCCTCGTGCACCAGCTTCTCGTGGACTCC
35 PDQGATDALHAEHVVEEHLR

CTGCTCGATCAGGAGCCGGCCCCTGACCGGATCGGTGTGGACGTCTCCCGGATCGGCCTC

56200 + + + + + + 56259

GACGAGCTAGTCCT CGGCCGGGGACTGGC CTAGCCACAC CTGCAGAGGGCCTAGCCGGAG
35 LLDQEPAPDRIGVDVSRIGL

GCCCACTCGGCGCAGACTGCCCTGCGCAACACCGGGCGGCTGGCTGCCGCTGCGGCGCTC

56260 + + + + + + 56319

CGGGTGAGCCGCGTCTGACGGGACGCGTTGTGGCCCGCCGACCGACGGCGACGCCGCGAG

70

A

HSAQTALRNTGRLAAAAAL

CGCGGGAGCGAGGTCGTCCTGCTCATGGAGGGGTCCGAGGACATCGACACCGTGCTGGCC

56320 + + + + + + 56379

GCGCCCTCGCTCCAGCAGGACGAGTACCTCCCCAGGCTCCTGTAGCTGTGGCACGACCGG

RGSEVVLLMEGSEDIDTVLA

GTCCATGACGCCCTGGTGAACCGTTACGACAACGTGGGGATCACCCTTCAGGCGCACCTG

56380 + + + + + + 56439

CAGGTACTGCGGGACCACTTGGCAATGCTGTTGCACCCCTAGTGGGAAGTCCGCGTGGAC

VHDALVNRYDNVG I TLQAHL

CACCGCACCGTGGACGACGCCATGGCGGTCGCGGGTCCTGGCCGCACCGTGCGGCTGGTC

56440 + + + + + + 56499

GTGGCGTGGCACCTGCTGCGGTACCGCCAGCGCCCAGGACCGGCGTGGCACGCCGACCAG

H R

TVDDAMAVAGPGRTVRLV

ATGGGCTCCTCGGCCGAGCCTGCCGGCACCGCTCTGTCCCGGGGCCCCGCTCTGGAGGAC

56500 + + + + + + 56559

TACCCGAGGAGCCGGCTCGGACGGCCGTGGCGAGACAGGGCCCCGGGGCGAGACCTCCTG

MGSSAEPAGTALSRGPALED

CGGTACCTTGACCTCGCGGAGCTTCTCGTGGACCGTGGCGTCCGGCTGAGTCTGGCCACT

56560 + + + + + + 56619

GCCATGGAACTGGAGCGCCTCGAAGAGCACCTGGCACCGCAGGCCGACTCAGACCGGTGA

RYLDLAELLVDRGVRLSLAT

CCGGACGCCGAGGTCCTGGCCGGGGCGCAGGAGCGTGGTCTGCTCGAACGCGTCCAGGAC

56620 + + + + + + 56679

GGGCTGCGGCTCCAGGACCGGCCCCGCGTCCTCGCACCAGACGAGCTTGCGCAGGTCCTG
PDAEVLAGAQERGLLERVQD

ATCGAGATGCTCTACGGTGTGCGGCCCGAGCTGCTGCGCCGCCACCGGGCGGCGGGCCGC

56680 + + + + + + 56739

TAGCTCTACGAGATGCCACACGCCGGGCTCGACGACGCGGCGGTGGCCCGCCGCCCGGCG
IEMLYGVRPELLRRHRAAGR

CCCTGTCGCATCCACGCGGCCTACGGGATGAACTGGTGGCTTCCCCTGCTGCGGAGGCTG

56740 + + + + + + 56799

GGGACAGCGTAGGTGCGCCGGATGCCCTACTTGACCACCGAAGGGGACGACGCCTCCGAC

PCRIHAAYGMNWWLPLLRRL

GCCGACAACCCGCCGATGGTGCTCAACGCCCTGGCCGACATCGGCCGGGACCGGGAGCCC

56800 + + + + + + 56859

CGGCTGTTGGGCGGCTACCACGAGTTGCGGGACCGGCTGTAGCCGGCCCTGGCCCTCGGG
ADNPPMVLNALADIGRDREP

GTCGCCCACCAGGCGTACTGACCCGCCCCGGGCCGCGATCCGCGGGGCACCGGCCCCGGG

56860 + + + + + + 56919

CAGCGGGTGGTCCGCATGACTGGGCGGGGCCCGGCGCTAGGCGCCCCGTGGCCGGGGCCC

V A H Q A Y * (OF35)

GCGCCGGTCAGCTCCCGGTCGCCGCGAACTGCCCGGGCCTGCGCCCCTCGCCCGCCGGCC

56920 + + + + + + 56979

CGCGGCCAGTCGAGGGCCAGCGGCGCTTGACGGGCCCGGACGCGGGGAGCGGGCGGCCGG
(ORF36) * SGTAAFQGPRRGEGAP

CCCGGTAGGCCTGGGCGATGTCCAGCCACTTCTCCGCCTCCTGACCAGACGCGGTCAGGG

56980 + + + + + + "039

GGGCCATCCGGACCCGCTACAGGTCGGTGAAGAGGCGGAGGACTGGTCTGCGCCAGTCCC
; GRYAQAIDLWKEAEQGSATL

CGAGGTCGTCGCGGTGGCGGCGCCGGGTGACCAGCAGGCAGAAGTCGTGCGCGGGACCGC

57040 + + + + + + 57099

GCTCCAGCAGCGCCACCGCCGCGGCCCACTGGTCGTCCGTCTTCAGCACGCGCCCTGGCG
; ALDDRHRRRTVLLCFDHAPG

71

TGACCGTCTCGGTGGCGTCCTCGGGGCCGACCGTCCAGACCTCGCCCGAGGGGGCGGTGA

57100 + + + + + + 57159

ACTGGCAGAGCCACCGCAGGAGCCCCGGCTGGCAGGTCTGGAGCGGGCTCCCCCGCCACT
36 SVTETADEPGVTWVEGS PAT

GCTCGAAGCGGAACGGCGCGGCCGGCGGGGTCAGACCGTGGGACTCGTAGCCGAAGTCGC

57160 + + + + + + 57219

CGAGCTTCGCCTTGCCGCGCCGGCCGCCCCAGTCTGGCACCCTGAGCATCGGCTTCAGCG
36 LEPRFPAAPPTLGHSEYGFD

GTGTCAGCCAGGCGAAGTCGACGATGTTGCGAAGCCGCTCGGTGGGCGTGCGCCGGACAC

57220 + + + + + + 57279

CACAGTCGGTCCGCTTCAGCTGCTACAACGCTTCGGCGAGCCACCCGCACGCGGCCTGTG
36 RTLWAFDVINRLRETPTRRV

CCAGGGCGTCGGCGACGTCCTGGCCGTGGGCGAACACCTCCATGATCCCGGCGCAGCCCA

57280 + + + + + + 57339

GGTCCCGCAGCCGCTGCAGGACCGGCACCCGCTTGTGGAGGTACTAGGGCCGCGTCGGGT
36 GLADAVDQGHAFVEMI GACG

GAACGACCGGCGGCAGCGGGTTGACCAGCCACGGAACCACCTGGCCGGCGGGGACCGCGG

57340 + + + + + + 57399

CTTGCTGGCCGCCGTCGCCCAACTGGTCGGTGCCTTGGTGGACCGGCCGCCCCTGGCGCC
36 LVVPPLPNVLWPVVQGAPVA

CGAGCGCCTCGACCGAGGCCCGCCCCATGCCCCGGAAGCGGGTGAGCAGTTCCTGCGGCG

57400 + + + + + + 57459

GCTCGCGGAGCTGGCTCCGGGCGGGGTACGGGGCCTTCGCCCACTCGTCAAGGACGCCGC
36 ALAEVSARGMGRFRTLLEQP

GGAAGCCCTTGAACTGCTGCAGAGCCGCGTTGACCGCTCCGTCGAAGTTGCCTGCCGCGG

57460 + + + + + + 57519

CCTTCGGGAACTTGACGACGTCTCGGCGCAACTGGCGAGGCAGCTTCAACGGACGGCGCC
36 PFGKFQQLAANVAGD FNGAA

CGGCCGTGACGGCCTTGAACTCCTCCGGCGCCGCCGCCGCGGTCCTGGCCAGGTTGAAGA

57520 + + + + + + 57579

GCCGGCACTGCCGGAACTTGAGGAGGCCGCGGCGGCGGCGCCAGGACCGGTCCAACTTCT
36 AATVAKFEE PAAAATRALNF

CGAAGGTGAGGTGGGCGATCTGGTCGGTGACGGTCCAGCCGGGCGCCGGCGTCGGAGTGT

57580 + + + + + + 57639

GCTTCCACTCCACCCGCTAGACCAGCCACTGCCAGGTCGGCCCGCGGCCGCAGCCTCACA
36 VFTLHAIQDTVTWGPAPTPT

TCCAGGCTTCGTCGTCGATCTTCTCGACCAGCTGCGCCAGCTCCTCGATGTCGGTGGCCA

57640 + + + + + + 57699

AGGTCCGAAGCAGCAGCTAGAAGAGCTGGTCGACGCGGTCGAGGAGCTACAGCCACCGGT
36 NWAEDDIKEVLQALEEIDTA

GGTGCTTGAGGACGTCGTCGAGCGAATTCATCTCGTACTTCCTTCACTGGGGGTGTTCCG

57700 + + + + + + 57759

C CACG7VACTCCTGCAGCAGCTCGCTTAAGTAGAGCATGAAGGAAGTGACC CCCACAAGGC
36 LHKLVDDLSNM (ORF36)

GGCTGGGACGGATGTCCCGCCGGGTGGGCCGGCGGCCGGCGGAAGCGCCGTCGCGGAGCG

57760 + + + + + + 57819

CCGACCCTGCCTACAGGGCGGCCCACCCGGCCGCCGGCCGCCTTCGCGGCAGCGCCTCGC

TCGGCGACAGTCGCTAGGCGGCGCGTCCCGCGTAGGAGCCGGCCCGGTCGGAATAGGGCG

57820 + + + + + + 57879

AGCCGCTGTCAGCGATCCGCCGCGCAGGGCGCATCCTCGGCCGGGCCAGCCTTATCCCGC
37 (ORF37) *AARGAYSGARDSYP

CGAGCGCCTCGGCCAGGGCTTCGGGTATCAGGGTCGGCACGGTCGCCGTGTTGGGGCCGC

72

57880 + + + + + + 57939

GCTCGCGGAGCCGGTCCCGAAGCCCATAGTCCCAGCCGTGCCAGCGGCACAACCCCGGCG
37 ALAEALAEPILTPVTATNPG

GCATGCAGGCGATGCGCTGGCGTCCCCGCGCCACCAGGGTCTCGCCGCCGTCGTCGCCCA

57940 + + + + + + 57999

CGTACGTCCGCTACGCGACCGCAGGGGCGCGGTGGTCCCAGAGCGGCGGCAGCAGCGGGT
37 RMCAIRQRGRAVLTEGGDDG

GCTTGATGTAGTCGAAGGTGAACTCCAGCTGGGTCTGCCGCAGCTCCGAGAGCCTCATCC

58000 + + + + + + 58059

CGAACTACATCAGCTTCCACTTGAGGTCGACCCAGACGGCGTCGAGGCTCTCGGAGTAGG
37 LKIYDFTFELQTQRLESLRM

GGATCGACAGTTCGTCGAAGGCGGTGATCTCCGCGAAGAACTCGCAGTCCACCTTGAGGG

58060 + + + + + + 58119

CCTAGCTGTCAAGCAGCTTCCGCCACTAGAGGCGCTTCTTGAGCGTCAGGTGGAACTCCC
37 RISLEDFATIEAFFECDVKL

TGAAGAGCTTGAGGTCCTCCTGGACCTCGGCGAGCACCGAAGGCGCCCTCTCCTTGAGAA

58120 + + + + + + 58179

ACTTCTCGAACTCCAGGAGGACCTGGAGCCGCTCGTGGCTTCCGCGGGAGAGGAACTCTT
37 TFLKLDEQVEALVSPAREKL

AGAGTTCCCGGCAACGCCCCTGCCAACGAAGGTAGTTGACGTAGTAGACGTTGCCGACGA

58180 + + + + + + 58239

TCTCAAGGGCCGTTGCGGGGACGGTTGCTTCCATCAACTGCATCATCTGCAACGGCTGCT
37 FLERCRGQWRLYNVYYVNGV

GGTTCGTCTCCTCGAAGCCGACGGTGTGGCGGAGCTCGAAGTAGTCAGGATTCGTCGCGG

58240 + + --- + + + + 58299

CCAAGCAGAGGAGCTTCGGCTGCCACACCGCCTCGAGCTTCATCAGTCCTAAGCAGCGCC
37 LNTEEFGVTHRLEFYDPNTA

TCATAGGTCTGTGCCCTTCGTCGTCGGGGCCGGTCGTCGCACCGAGTTGCGTGAAGCAAC

58300 + + + + + + 58359

AGTATCCAGACACGGGAAGCAGCAGCCCCGGCCAGCAGCGTGGCTCAACGCACTTCGTTG

37 T M (ORF37)

TCACTGGTCGCGATGGCCTGCGGGGTCGGTGGCCCGCGCTCCGGGCGGAGAGTGCGGGCG

58360 + + + + + + 58419

AGTGACCAGCGCTACCGGACGCCCCAGCCACCGGGCGCGAGGCCCGCCTCTCACGCCCGC

GGGTGCCGGCCGGCGCGGGGTCAGCCGCGCGCCGACGGCAGCAGGGGAAGAACCCTCTCG

58420 + + + + + + 58479

CCCACGGCCGGCCGCGCCCCAGTCGGCGCGCGGCTGCCGTCGTCCCCTTCTTGGGAGAGC

38 (ORF38) *GRAS PLLPLVRE

CGGCCGCTCGTGGAGCCGTCGGGGGCCGGTGCGCCGTAGGTGACGGAGATACCCCGGCTC

58480 + + + + + + 58539

GCCGGCGAGCACCTCGGCAGCCCCCGGCCACGCGGCATCCACTGCCTCTATGGGGCCGAG
38 RGSTSGDPAPAGYTVS I GRS

TGCGCGGCGCGCACGATCCCCGGCATCGCGCGTTCGGCGAGCGCCGCGATGGTCATCGCG

58540 + + + + + + 58599

ACGCGCCGCGCGTGCTAGGGGCCGTAGCGCGCAAGCCGCTCGCGGCGCTACCAGTAGCGC
38 QAARVIGPMAREALAAITMA-

GGATTGACCGTCAGCGCGCCGGGAACCGACGATCCGTCGGTGACGAAGATCCCCGGGTGG

58600 + + + + + + 58659

CCTAACTGGCAGTCGCGCGGCCCTTGGCTGCTAGGCAGCCACTGCTTCTAGGGGCCCACC
38 PNVTLAGPVSSGDTVFIGPH

TCGCGGAGCTCGTTGCTGTCGTCCAGGGCGGATGTGTGGGGGTCGTCGCCCATCCGGCAG

58660 + + + + + + 58719

AGCGCCTCGAGCAACGACAGCAGGTCCCGCCTACACACCCCCAGCAGCGGGTAGGCCGTC

73

38 DRLENSDDLASTHPDDGMRC

GAGGAGAGCGGGTGGACGGTGTAGGCGCCGACGAGGTCGTTGGTCCAGGGCATGACCTTG

58720 + + + + + + 58779

CTCCTCTCGCCCACCTGCCACATCCGCGGCTGCTCCAGCAACCAGGTCCCGTACTGGAAC
38 SSLPHVTYAGVLDNTWPMVK-

GCCAGGCCGTCCTTCTCCAGGATCTCCTTGACCTCGGCGTCGGATGCGGCCCAGGCGCCC

58780 + + + + + + 58839

CGGTCCGGCAGGAAGAGGTCCTAGAGGAACTGGAGCCGCAGCCTACGCCGGGTCCGCGGG
38 ALGDKEL I EKVEADSAAWAG

AGGGTGTTCTTCGTCGGGTCGTAGCGCAGGTTGCCCCGGCCGAGCATCTGCTGGGAGATG

58840 + + + + + + 58899

TCCCACAAGAAGCAGCCCAGCATCGCGTCCAACGGGGCCGGCTCGTAGACGACCCTCTAC
38 LTNKTPDYRLNGRGLMQQS I

CGGTGGGCGTTACCGGTGGCGGGAGGGGGGCCGAAGACGCCTTCGTTGTCGTCCTCGATC

58900 + + + + + + 58959

GCCACCCGCAATGGCCACCGCCCTCCCCCCGGCTTCTGCGGAAGCAACAGCAGGAGCTAG
38 RHANGTAPPPGFVGENDDEI-

ATCGTGAAGATCGTGAGCCAGGAGGTCCACTGCTTCAGGATCTCCTTCTTCTCCTTGCCG

58960 + + + + + + 59019

TAGCACTTCTAGCACT CGGTCCTC CAGGTGACGAAGTCCTAGAGGAAGAAGAGGAACGGC
38 MTFITLWSTWQKLIEKKEKG-

AACCAGGAGGGGCCCGTGGCGCCGGGCACCTGGGCGAGGATCGTGCCGAGGCCCGGCGGG

59020 + + + + + + 59079

TTGGTCCTCCCCGGGCACCGCGGCCCGTGGACCCGCTCCTAGCACGGCTCCGGGCCGCCC
38 FWSPGTAGPVQALITGLGPP

AAGTAGAGCTGTTCCAGGGAGTAGCGGGAGTACTCGGGCAACGAGCCGTCCAGCCTGTCC

59080 + + + + + + 59139

TTCATCTCGACAAGGTCCCTCATCGCCCTCATGAGCCCGTTGCTCGGCAGGTCGGACAGG
38 FYLQELSYRSYEPLSGDLRD-

CAGCTCGCCACGGTGGGCCCCTTGCCGATCTGGTTGGCCGCGTAGGCGAGCCCGTCGCCC

59140 + + + + + + 59199

GTCGAGCGGTGCCACCCGGGGAACGGCTAGACCAACCGGCGCATCCGCTCGGGCAGCGGG
38 WSAVTPGKGIQNAAYALGDG-

CGGTCCAGGCCGAACAGCTCGGCCGCCTTGGCCTCGTCGATGATGGCGGTGTTGAGCCGC

59200 + + + + + + 59259

GCCAGGTCCGGCTTGTCGAGCCGGCGGAACCGGAGCAGCTACTACCGCCACAACTCGGCG
38 RDLGFLEAAKAEDI IATNLR-

TCGCCGTTGCCGGAGAAGTAGCGTCCGACCGCTCGTGGCATGGTGCCCAGGTGGGCCTCG

59260 + + + + + + 59319

AGCGGCAACGGCCTCTTCATCGCAGGCTGGCGAGCACCGTACCACGGGTCCACCCGGAGC
38 EGNGS FYRGVARPMTGLHAE

CTGCGCTGGAGGATCACCGGGGTCGCGCCCGCGCCGGCCGCCATCACCACGATCTTCGCC

59320 + + + + + + 59379

GACGCGACCTCCTAGTGGCCCCAGCGCGGGCGCGGCCGGCGGTAGTGGTGCTAGAAGCGG
38 SRQL IVPTAGAGAAMVVI KA-

TCGATGACGCCGCTGCCCGCCTGGAGGCGGTAGTCGTCGTCGTGCACGACGTTGTAGTGC

59380 + + + + + + 59439

AGCTACTGCGGCGACGGGCGGACCTCCGCCATCAGCAGCAGCACGTGCTGCAACATCACG
38 E IVGS GAQLRYDDDHVVNYH

ACCCGGTAGGAGCCGTCGGGGGTGCGCGAGAGGTGCTGGACCTCGTGCAGCGGGCGGATG

59440 + + + + + + 59499

TGGGCCATCCTCGGCAGCCCCCACGCGCTCTCCACGACCTGGAGCACGTCGCCCGCCTAC
38 VRYSGDPTRSLHQVEHLPRI-

74

CGCGCCCCATGGGCGATGGCGGCGGGCAGGTAGTTGACCAGCAAGGACTGCTTGGCCTCG

59500 + + + + + + 59559

GCGCGGGGTACCCGCTACCGCCGCCCGTCCATCAACTGGTCGTTCCTGACGAACCGGAGC
38 RAGHAIAAPLYNVLLSQKAE

AAGCGGCAGCCGGCCATCATCCAGTTGCAGTTCACGCACTTGGTGTTGTCGATGGCGACG

59560 + + + + + + 59619

TTCGCCGTCGGCCGGTAGTAGGTCAACGTCAAGTGCGTGAACCACAACAGCTACCGCTGC
38 FRCGAMMWNCNVCKTNDIAV-

GCGAGGGGGTTGGCGGTGCGGCCGGCGTGGTTGCACGCCGCGGCCCACAGTCCGCCGGCG

59620 + + + + + + 59679

CGCTCCCCCAACCGCCACGCCGGCCGCACCAACGTGCGGCGCCGGGTGTCAGGCGGCCGC
38 ALPNATRGAHNCAAAWLGGA-

TAGCTCACGTCGTTCCAGTCCTGCCGGGTCACGGAGAGGGACTCCTCGACACGGTCGTAC

59680 + + + + + + 59739

ATCGAGTGCAGCAAGGTCAGGACGGCCCAGTGCCTCTCCCTGAGGAGCTGTGCCAGCATG
38 YSVDNWDQRTVSLSEEVRDY-

CAGGGGTCCAGGGTTTCGCGGCTCACCGCCTGCGGCCACATCCGGCGTCCTATGGACCCC

59740 + + + + + + 59799

GTCCCCAGGTCCCAAAGCGCCGAGTGGCGGACGCCGGTGTAGGCCGCAGGATACCTGGGG
38 WPDLTERSVAQPWMRRGI SG

TGCCGGTCGAAGACGAAGCGCGGGGCGCGGGGCATCGCGGCGAAGTAGACGACGCTGCCG

59800 + + + + + + 59859

ACGGCCAGCTTCTGCTTCGCGCCCCGCGCCCCGTAGCGCCGCTTCATCTGCTGCGACGGC
38 QRDFVFRPARPMAAFYVVSG-

CCGCCCACACAGTTCCCGCCGAGGATGCTCATGCCGTCCCCGACCGTGAAGTCGAACGCC

59860 + + + + + + 59919

GGCGGGTGTGTCAAGGGCGGCTCCTACGAGTACGGCAGGGGCTGGCACTTCAGCTTGCGG
38 GGVCNGGLI SMGDGVTFDFA-

CTCGTGTACGAGGAGCCGAGTTTGTAGTCGTGCTCGAACTCCTTGCTCTCCAGCCACGGC

59920 + + + + + + 59979

GAGCACATGCTCCTCGGCTCAAACATCAGCACGAGCTTGAGGAACGAGAGGTCGGTGCCG
38 RTYSSGLKYDHEFEKSELWP

CCGCGTTCCAGGACGGTGACGTCGGCGCCCCCCGCCGCCAGGTGGTAGGCGGCGATGGCA

59980 + + + + + + 60039

GGCGCAAGGTCCTGCCACTGCAGCCGCGGGGGGCGGCGGTCCACCATCCGCCGCTACCGT
38 GRELVTVDAGGAALHYAAIA-

CCGCCGAATCCGCTGCCGATGACGAGGACGTCCGTGCGCTCGGCCGTGGTGCTCATGCGG

60040 + + + + + + 60099

GGCGGCTTAGGCGACGGCTACTGCTCCTGCAGGCACGCGAGCCGGCACCACGAGTACGCC

38 GGFGSGIVLVDTREATTSM

(ORF3 9) * A

GGCTCCCGGTGGACGTGGTGTCGGGGTGGAGGCGGGCGAACTCACGCCCGTAGCTGTAAT

60100 + + + + + + 60159

CCGAGGGCCACCTGCACCACAGCCCCACCTCCGCCCGCTTGAGTGCGGGCATCGACATTA

39 PSGTSTTDPHLRAFERGYSY

CCTTGAAGCGCCACAGGCCGTCGGCGTCCGGCATGCTCAGGCCCATGGCCTCCAGTCCCG

60160 + + + + + + 60219

GGAACTTCGCGGTGTCCGGCAGCCGCAGGCCGTACGAGTCCGGGTACCGGAGGTCAGGGC
39 DKFRWLGDADPMSLGMAELG

GATGGCCGTCCTCCATCGCCTGTGCCGTGTTGAGGTGCGCGGCCGAATCGAAGGCCATGT

60220 + + + + + + 60279

CTACCGGCAGGAGGTAGCGGACACGGCACAACTCCACGCGCCGGCTTAGCTTCCGGTACA
39 PHGDEMAQATNLHAASDFAM

75

TGCAGAAGAGGGACAGCAGCACCCAGAACTCCTTCTCGGGGTGGCCTGGTGTCGTCAGCC

60280 + + + + + + 60339

ACGTCTTCTCCCTGTCGTCGTGGGTCTTGAGGAAGAGCCCCACCGGACCACAGCAGTCGG
39 NCFLSIrLVWFEKEPHGPTTL

GCTGGATCAGCGCGGCCCGGTCCGGGTAGTCGAGCGCCACGAAGGGCGGGACCGTCGGGT

60340 + -f + + + + 60399

CGACCTAGTCGCGCCGGGCCAGGCCCATCAGCTCGCGGTGCTTCCCGCCCTGGCAGCCCA
39 RQILAARDPYDLAVFPPVTP

CGGGAGCCAGGCGGCGCTCCGCCGCGTAGGCCAGCGCGTGCTCGTTCACCAGGCGCACCA

60400 + + + + + + 60459

GCCCTCGGTCCGCCGCGAGGCGGCGCATCCGGTCGCGCACGAGCAAGTGGTCCGCGTGGT
39 DPALRREAAYALAHENVLRV

GGTCGTCCAGACCCTCGTGGATGCCGGTCGCATCCCATTGCAGGAGCTCCAGGGCTCCCG

60460 + + -f + + + 60519

CCAGCAGGTCTGGGAGCACCTACGGCCAGCGTAGGGTAACGTCCTCGAGGTCCCGAGGGC
39 LDDLGEHIGTADWQLLELAG

CCTGGACGGCGCCACCGCCGGTGGACACCCCCGCGATGGCCCGGTCGTCCGCGAAGCGCT

60520 + + + + + + 60579

GGACCTGCCGCGGTGGCGGCCACCTGTGGGGGCGCTACCGGGCCAGCAGGCGCTTCGCGA
39 AQVAGGGTSVGAIARDDAFR

TCTGGCCCGGCACGATCGTGTCCGCGTAGGCCTCCAGGGTCATGGTCCGGATATCGCCGG

60580 + + + + + + 60639

AGACCGGGCCGTGCTAGCACAGGCGCATCCGGAGGTCCCAGTACCAGGCCTATAGCGGCC

39 KQGPVITDAYAELTM (ORF3 9)

CCGGCGCCCCTCGCTCATTGTCGTCGCGCAACTCGCTCTCCATTCTCGCAGTCCGGAGTG

60640 + + + + + + 60699

GGCCGCGGGGAGCGAGTAACAGCAGCGCGTTGAGCGAGAGGTAAGAGCGTCAGGCCTCAC

GGATGCCTTGTGGCGAGGAGAAAGCTAGGTTCGTTCGACCGGTTCAAGCAACTAGCCAAA

60700 + + + + + + 60759

CCTACGGAACACCGCTCCTCTTTCGATCCAAGCAAGCTGGCCAAGTTCGTTGATCGGTTT

GTCGAGGCGACCTTGAAACCGACTCCACGGAGTTGGCGCGAAGCGGCGGATGGATTACAC

60760 + + + + + + 60819

CAGCTCCGCTGGAACTTTGGCTGAGGTGCCTCAACCGCGCTTCGCCGCCTACCTAATGTG

GCGCGGGCGAGCGGCTCACTAGTCTGGCCGCACGGATGTCTTCATCACCTGCACGTGGAA

60820 + + + + + + 60879

CGCGCCCGCTCGCCGAGTGATCAGACCGGCGTGCCTACAGAAGTAGTGGACGTGCACCTT

AAGCTTCTGCACGGGCACCGCATGTGGAAGTGAGCCCTGGTCTCATGTCTTGGGGGAAAC

60880 + + + + + + 60939

TTCGAAGACGTGCCCGTGGCGTACACCTTCACTCGGGACCAGAGTACAGAACCCCCTTTG

GTGAAAAGTGACTCTGCCCAACGCGCCGTGGAGCGATCACGCCGTGTCGTACGGATCGAT

60940 + + + + + + 60999

CACTTTTCACTGAGACGGGTTGCGCGGCACCTCGCTAGTGCGGCACAGCATGCCTAGCTA

40 VKSDSAQRAVERSRRVVRID

(ORF40)

GAACTCATTCCCGCCGATTCCCCGCGCCTGAACGGAATCGATCGTTCCCATGTGCAGCGC

61000 + + + + + + 61059

CTTGAGTAAGGGCGGCTAAGGGGCGCGGACTTGCCTTAGCTAGCAAGGGTACACGTCGCG
40 ELI PADS PRLNGIDRSHVQR

CTCGCGACCGTGTACGCGTCCCTGCCGCCGGTCCTGGTGCACCGCCCGACCATGCGGGTC

61060 + + + + + + 61119

GAGCGCTGGCACATGCGCAGGGACGGCGGCCAGGACCACGTGGCGGGCTGGTACGCCCAG
40 LATVYASLPPVLVHRPTMRV

76

GTCGACGGCATGCACCGCATCGGCGCGGCCCGCCTGAAGGGGCTGGACACGGTCGAGGTC

61120 + + + + + + 61179

CAGCTGCCGTACGTGGCGTAGCCGCGCCGGGCGGACTTCCCCGACCTGTGCCAGCTCCAG
40 VDGMHRIGAARLKGLDTVEV

ACCTTCTTCGAGGGCGCCGAGGAGCAGGTGTTCCTGCGTTCCGTCGCGGCGAACATCACC

61180 + + + + + + 61239

TGGAAGAAGCTCCCGCGGCTCCTCGTCCACAAGGACGCAAGGCAGCGCCGCTTGTAGTGG
40 TFFEGAEEQVFLRSVAANIT

AACGGCCTGCCGTTGTCGGTGGCCGACCGCAAGACCGCCGCGGCCCGCATTCTGGCCTCC

61240 + + + + + + 61299

TTGCCGGACGGCAACAGCCACCGGCTGGCGTTCTGGCGGCGCCGGGCGTAAGACCGGAGG
40 NGLPLSVADRKTAAARI LAS

CACCCGACCCTGTCCGACCGCGCGGTCGCCGCACACGTCGGCCTCGACGCCAAGACCGTG

61300 + + + + + + 61359

GTGGGCTGGGACAGGCTGGCGCGCCAGCGGCGTGTGCAGCCGGAGCTGCGGTTCTGGCAC
40 HPTLSDRAVAAHVGLDAKTV

GCGGGGGTACGGACGTGTTCAGCCGCGGGTTCTCCGCTGCTGAACATGCGCACCGGGGCG

61360 + + + + + + 61419

CGCCCCCATGCCTGCACAAGTCGGCGCCCAAGAGGCGACGACTTGTACGCGTGGCCCCGC
40 AGVRTCSAAGS PLLNMRTGA

GACGGCCGCGTCCACCCGTTGGACCGCACCGCCGAACGCCTGCACGCGGCCGCGCTGCTG

61420 + + + + + + 61479

CTGCCGGCGCAGGTGGGCAACCTGGCGTGGCGGCTTGCGGACGTGCGCCGGCGCGACGAC
40 DGRVHPLDRTAERLHAAALL

ACCCAGGACCCGGGACTCCCGTTGCGCTCCGTCGTCGAGCAGACGGGGCTGTCGCTGGGC

61480 + + + + + + 61539

TGGGTCCTGGGCCCTGAGGGCAACGCGAGGCAGCAGCTCGTCTGCCCCGACAGCGACCCG
40 TQDPGLPLRSVVEQTGLSLG

ACGGCCCACGACGTCCGCCGTCGGCTGCTGCGGGGCGAGGACCCGGTCCCGCAGAACCGG

61540 + + + + + + 61599

TGCCGGGTGCTGCAGGCGGCAGCCGACGACGCCCCGCTCCTGGGCCAGGGCGTCTTGGCC
40 TAHDVRRRLLRGED PVPQNR

CAGAGCGCGATGCTGGAGCCGGGACTCGCCCCGCAGAAGAAGGCGACGGCCAAGCCGCCC

61600 + + + + + + 61659

GTCTCGCGCTACGACCTCGGCCCTGAGCGGGGCGTCTTCTTCCGCTGCCGGTTCGGCGGG
40 QSAMLEPGLAPQKKATAKPP

GTCGGCCCGGCCGCCCGTCCGGTCCCGAAGGTGCCGCCCGCCGTCGCCGGCAGGCCGCCG

61660 + + + + + + 61719

CAGCCGGGCCGGCGGGCAGGCCAGGGCTTCCACGGCGGGCGGCAGCGGCCGTCCGGCGGC
40 VGPAARPVPKVPPAVAGRPP

GTGTCACCGCGGTCCCGGGCCCCGCTGGAGGCGCTGCGCAAGCTCTCCAACGACCCCTCC

61720 + + + + + + 61779

CACAGTGGCGCCAGGGCCCGGGGCGACCTCCGCGACGCGTTCGAGAGGTTGCTGGGGAGG
40 VSPRSRAPLEALRKLSNDPS

CTGCGCCACTCCGACCAGGGGCGCGAACTCATGCGCTGGCTGCACAACCGGTTCGTCGTC

61780 + + + + + + 61839

GACGCGGTGAGGCTGGTCCCCGCGCTTGAGTACGCGACCGACGTGTTGGCCAAGCAGCAG
40 LRHSDQGRELMRWLHNRFVV

GACGAGGCGTGGCGCCGGCGCGCGGACGCGGTCCCGGCCCACTGCGTCGACTCGATGGCG

61840 + + + + + + 61899

CTGCTCCGCACCGCGGCCGCGCGCCTGCGCCAGGGCCGGGTGACGCAGCTGAGCTACCGC
40 DEAWRRRADAVPAHCVDSMA

77

GAGCTGGCGCAGCACTGCTCGGACGCCTGGCACCGGTTCGCCGAGGAGATGGTTCGGCGC

61900 + + + + + + 61959

CTCGACCGCGTCGTGACGAGCCTGCGGACCGTGGCCAAGCGGCTCCTCTACCAAGCCGCG
40 ELAQHCSDAWHRFAEEMVRR

CGGCACAGCGCCGCGGCCGACGGCTCCGGACTCCGCACGACTCAGCCAACTCGCCGTTGA

61960 + + + + + + 62019

GCCGTGTCGCGGCGCCGGCTGCCGAGGCCTGAGGCGTGCTGAGTCGGTTGAGCGGCAACT

40 RHSAAADGSGLRTTQPTRR*
(ORF40) -

CGGCCTACTTCGACAGGGAGTTACGGTGACCACGAACACCATCGAGGACGCGGTCCGCCG

62020 + + + + + + 62079

GCCGGATGAAGCTGTCCCTCAATGCCACTGGTGCTTGTGGTAGCTCCTGCGCCAGGCGGC
41 (ORF41) VTTNT I EDAVRR

GGTCGTCGAGTACATGCACGTCAACCTGGGTCAGAACCTCACGATCGATGACATGGCGCG

62080 + + + + + + 62139

CCAGCAGCTCATGTACGTGCAGTTGGACCCAGTCTTGGAGTGCTAGCTACTGTACCGCGC
41 VVEYMHVNLGQNLT I DDMAR

CACGGCGATGTTCAGCAAGTTCCATTTCACCCGCATCTTCCGCGAAGTCACCGGTACCTC

62140 + + + + + + 62199

GTGCCGCTACAAGTCGTTCAAGGTAAAGTGGGCGTAGAAGGCGCTTCAGTGGCCATGGAG

41 TAMFSKFHFTRI FREVTGTS

TCCCGGGCGTTTCCTGTCCGCCTTACGGATTCAGGAGGCCAAGAGACTTCTCGTGCACAC

62200 + + + + + + 62259

AGGGCCCGCAAAGGACAGGCGGAATGCCTAAGTCCTCCGGTTCTCTGAAGAGCACGTGTG
41 PGRFLSALRI QEAKRLLVHT

TGCACTCAGTGTGGCCGATATCAGCAGTCAGGTCGGCTACAGCAGTGTCGGTACTTTCAG

62260 + + + + + + 62319

ACGTGAGTCACACCGGCTATAGTCGTCAGTCCAGCCGATGTCGTCACAGCCATGAAAGTC
41 ALSVADISSQVGYSSVGTFS-

TTCTCGCTTCAAGGCCTGTGTGGGGCTTTCCCCGAGCGCCTATCGCGACTTCGGCGGGGT

62320 + + + + + + 62379

AAGAGCGAAGTTCCGGACACACCCCGAAAGGGGCTCGCGGATAGCGCTGAAGCCGCCCCA
41 SRFKACVGLSPSAYRDFGGV-

GCAGCCGGGTTTTCCCTCCGCCGCGGCCCGTCTCACTCCCACCGCGCACAATCCCTCCGT

62380 + + + + + + 62439

CGTCGGCCCAAAAGGGAGGCGGCGCCGGGCAGAGTGAGGGTGGCGCGTGTTAGGGAGGCA
41 QPGFPSAAARLTPTAHNPSV-

GCGCGGCCGCATTCACTCCGCCCCGGGTGACAGGCCCGGAAGGATCTTCGTGGGCCTGTT

62440 + + + + + + 62499

CGCGCCGGCGTAAGTGAGGCGGGGCCCACTGTCCGGGCCTTCCTAGAAGCACCCGGACAA
41 RGRIHSAPGDRPGRI FVGLF-

CCCCGGCAGGATGCGCCAGGGCCGCCCGGCGCGCTGGACCGTCATGGAGAGTCCCGGGGC

62500 + + + + + + 62559

GGGGCCGTCCTACGCGGTCCCGGCGGGCCGCGCGACCTGGCAGTACCTCTCAGGGCCCCG
41 PGRMRQGRPARWTVMES PGA-

CTTCGAGCTCCGGGACGTGCCCGTGGGCACCTGGCACATCCTGGTCCACTCCTTCCCCGC

62560 + + + + + + 62619

GAAGCTCGAGGCCCTGCACGGGCAC CCGTGGACCGTGTAGGAC CAGGTGAGGAAGGGGCG
41 FELRDVPVGTWHILVHSFPA-

CGGACACCGGCCGCACCAGCTCGACTCCGAACCGCTGTTGCTCGGGCACAGCGGACCGCT

62620 + + + + + + 62679

GCCTGTGGCCGGCGTGGTCGAGCTGAGGCTTGGCGACAACGAGCCCGTGTCGCCTGGCGA
41 GHRPHQLDSEPLLLGHSGPL-

78

CGTGGTGCACCCCGGTGCCCTGCTCCGGCCGGCGGACATCCTCCTGCGCGCGGTGGACGC

62680 + + + + + + 62739

GCACCACGTGGGGCCACGGGACGAGGCCGGCCGCCTGTAGGAGGACGCGCGCCACCTGCG
41 VVHPGALLRPADI LLRAVDA-

CCTCGATCCACCGGTCCTGCTGGCCCACTTCGCGCTGGAGAGCCGCCTCACCTCGCCGTA

62740 + + + + + + "799

GGAGCTAGGTGGCCAGGACGACCGGGTGAAGCGCGACCTCTCGGCGGAGTGGAGCGGCAT

41 LDPPVLLAHFALESRLTSPY-

42 (0RF42) * R A T -

CTCACCGTCATCGGTAGCCCTCCGCGCATCCGCAGGGAGAGCATGGGTTCGGCAACCGCC

62800 + + + + + + 62859

GAGTGGCAGTAGCCATCGGGAGGCGCGTAGGCGTCCCTCTCGTACCCAAGCCGTTGGCGG

41 SPSSVALRASAGRAWVRQPP-

42 SVTMPLGGRMRLSLMPEAVA-

CGGTGTCCGGCGACGGTACGCAGATCGAGATCGCGGGTGACCAGGGCCGTGACGAACACC

62860 + + + + + + 62919

GCCACAGGCCGCTGCCATGCGTCTAGCTCTAGCGCCCACTGGTCCCGGCACTGCTTGTGG

41 GVRRRYADRDRG* (ORF41)

42 RHGAVTRLDLDRTVLATVFV-

GCCTCCATCATCCCGAGGTTGCTGCCGACGCAGAACCGGGGCCCCGCGCCGAACGGGATG

62920 + + + + + + 62979

CGGAGGTAGTAGGGCTCCAACGACGGCTGCGTCTTGGCCCCGGGGCGCGGCTTGCCCTAC

42 AEMMGLNSGVCFRPGAGFPI-

TACGCGTACCGCGGCCGGTCGGCGGTCTGCCGGGGTTCGAACCGCTCGGGGTCGAAGCGC

62980 + + + + + + 63039

ATGCGCATGGCGCCGGCCAGCCGCCAGACGGCCCCAAGCTTGGCGAGCCCCAGCTTCGCG

42 YAYRPRDATQRPEFREPDFR-

TCGGGGTCCTCCCACAGCCCCGGATGGCGGTGCATGATGTACGGGCAGACCAGCACATCC

63040 + + + + + + 63099

AGCCCCAGGAGGGTGTCGGGGCCTACCGCCACGTACTACATGCCCGTCTGGTCGTGTAGG

42 EPDEWLGPHRHMIYPCVLVD-

42

GATCCGGCGGACACCGTGTAGCCGCCGACCACATCGCGTTGCTGGGCCACCCTGGGCAGG

+ + + + + +

CTAGGCCGCCTGTGGCACATCGGCGGCTGGTGTAGCGCAACGACCCGGTGGGACCCGTCC

SGASVTYGGVVDRQQAVRPL

ATCCC

63160 + 63164

TAGGG
42 I G -

79